B73410EN

Instructions for Use
DxFLEX
Flow Cytometer
C44966AB
Beckman Coulter Biotechnology (Suzhou) Co., Ltd.

Units 101, 201, 301, Building 4, No. 218 Sangtian Street,
BioBAY Sangtian Island, Suzhou Industrial Park,
Suzhou, P.R. China
DxFLEX
Flow Cytometer
PN C44966AB (August 2020)
© 2020 Beckman Coulter, Inc.

All rights reserved.
Trademarks
Beckman Coulter, the stylized logo, and the Beckman
Coulter product and service marks mentioned herein are
trademarks or registered trademarks of Beckman Coulter,
Inc. in the United States and other countries.
CytoFLEX and CytExpert are trademarks or registered
trademarks of Xitogen Technologies (Suzhou) Inc. in the
United States and other countries. Xitogen is a Beckman
Coulter company.
Windows 7, Windows 8, and Windows 10 are registered
trademarks or trademarks of Microsoft Corporation in the
United States and/or other countries.
All other trademarks are the property of their respective

owners.
Contact Us
If you have any questions, contact our Customer Support
Center.
• Worldwide, find us via our website at

www.beckman.com/support/technical
• In the USA and Canada, call us at 1-800-369-0333.
• Outside of the USA and Canada, contact your local
Beckman Coulter Representative.
Glossary of Symbols is available at
beckman.com/techdocs (PN C24689).
May be covered by one or more pat.- see

www.beckman.com/patents
Beckman Coulter Eurocenter S.A.

22, rue Juste-Olivier
Case Postale 1044
CH - 1260 Nyon 1, Switzerland
Tel: +41 (0) 22 365 36 11
Original Instructions
Revision History
Initial Issue AA, 11/2019
Software Version 2.0
Revision AB, 08/2020
Software Version 2.0
Links to the topics that changed are listed below:
CHAPTER 1, System Overview
Acoustic Noise Level
Electrical Ratings
Cytometer
CHAPTER 2, Using the CytExpert for DxFLEX Software
Start Page
Maintenance Log
Software Settings
CHAPTER 10, Troubleshooting
Backup/Restore
CHAPTER 12, Replacement/Adjustment Procedures
Changing Sample Mixing and Backflush Settings
APPENDIX D, Good Practices for Cyber Security
Drive Encryption
Protection from Malware Software
Operating System Updates
System Hardening
Remote Access
This document applies to the latest software listed and higher versions. When a subsequent software version affects the
information in this document, a new issue will be released to the Beckman Coulter Web site. For labeling updates, go to
www.beckman.com and download the latest version of the manual or system help for your instrument.
C44966AB iii
Revision History
iv C44966AB
Safety Notices
Read all product manuals and consult with Beckman Coulter-trained personnel before attempting
to operate instrument. Do not attempt to perform any procedure before carefully reading all
instructions. Always follow product labeling and manufacturer’s recommendations. If in doubt as
to how to proceed in any situation, contact us.
Beckman Coulter, Inc. urges its customers to comply with all national health and safety standards
such as the use of barrier protection. This may include, but it is not limited to, protective eyewear,
gloves, and suitable laboratory attire when operating or maintaining this or any other automated
laboratory analyzer.
This manual assumes that users have basic knowledge of the Windows operating system, as well as
experience working with laboratory testing technology. Users are invited to consult the
appropriate documentation for such information.
Alerts for Danger, Warning, and Caution
DANGER
DANGER indicates an imminently hazardous situation which, if not avoided, will
result in death or serious injury.
WARNING
WARNING indicates a potentially hazardous situation which, if not avoided, could
result in death or serious injury.
CAUTION
CAUTION indicates a potentially hazardous situation, which, if not avoided, may
result in minor or moderate injury. It may also be used to alert against unsafe
practices.
C44966AB v
Safety Notices
Safety Precautions
Safety Precautions
WARNING
Risk of operator injury if:
• All doors, covers and panels are not closed and secured in place prior to and
during instrument operation.
• The integrity of safety interlocks and sensors is compromised.
• Instrument alarms and error messages are not acknowledged and acted upon.
• You contact moving parts.
• You mishandle broken parts.
• Doors, covers and panels are not opened, closed, removed and/or replaced
with care.
• Improper tools are used for troubleshooting.
To avoid injury:
• Keep doors, covers and panels closed and secured in place while the
instrument is in use.
• Take full advantage of the safety features of the instrument.
• Acknowledge and act upon instrument alarms and error messages.
• Keep away from moving parts.
• Report any broken parts to your Beckman Coulter Representative.
• Open/remove and close/replace doors, covers and panels with care.
• Use the proper tools when troubleshooting.
CAUTION
System integrity could be compromised and operational failures could occur if:
• This equipment is used in a manner other than specified. Operate the
instrument as instructed in the product manuals.
• You introduce software that is not authorized by Beckman Coulter into your
computer. Only operate your system’s software with software authorized by
Beckman Coulter.
• You install software that is not an original copyrighted version. Only use
software that is an original copyrighted version to prevent virus
contamination.
CAUTION
If you purchased this product from anyone other than Beckman Coulter or an
authorized Beckman Coulter distributor, and, it is not presently under a Beckman
Coulter service maintenance agreement, Beckman Coulter cannot guarantee that
the product is fitted with the most current mandatory engineering revisions or
that you will receive the most current information bulletins concerning the
product. If you purchased this product from a third party and would like further
information concerning this topic, contact us.
vi C44966AB
Safety Notices
Symbol Explanations
CAUTION
Risk of instrument damage. This device is intended for indoor use only. To avoid
device damage, do not install the instrument outdoors.
WARNING
Risk of personal injury. Safety protection can be impaired if used in a manner not
specified by the manufacturer. To avoid personal injury, use the instrument
according to the manufacturer's instructions only.
Symbol Explanations
Symbol meanings
Consider all materials (specimens, reagents, controls, and monoclonal
antibodies) and areas these materials come into contact with as being
potentially infectious.
Wear appropriate barrier protection and follow safe laboratory procedures
when handling any material in the laboratory.
EMC Information
This in vitro diagnostic (IVD) equipment complies with the emission or anti-interference
requirement described in GB/T18268.26 (IEC 61326-2-6).
CAUTION
This equipment has been designed and tested to GB4824 (CISPR 11 Class A)
requirement. In a household environment, it could cause radio interference, in
which case, you may need to take measures to mitigate the interference.
It is advised that prior to operation of the device, the electromagnetism in the
environment should be evaluated. Do not use this device in close proximity to
sources of strong electromagnetic radiation (for example, unshielded RF sources),
as these could interfere with the proper operation of this device.
C44966AB vii
Safety Notices
EMC Information
viii C44966AB
Contents
Revision History, iii

Safety Notices, v
Alerts for Danger, Warning, and Caution, v
Safety Precautions, vi
Symbol Explanations, vii
EMC Information, vii
Introduction, xxv
Overview, xxv
How to Use Your Manual, xxv
About this Manual, xxv
Conventions Used, xxvii
Graphics, xxvii
CHAPTER 1: System Overview, 1-1

Overview, 1-1
Intended Use, 1-1
Main Components, 1-2
Optical Components, 1-3
Wavelength Division Multiplexer (WDM), 1-4
Optical Fiber, 1-7
Fluidics System, 1-8
Fluid Containers, 1-9
Fluidics Module, 1-10
Sample Station [Without Autoloader], 1-12
Sample Tube Holder Positions [Without Autoloader], 1-13
Sample Station [With Autoloader], 1-13
System Configuration [Without Autoloader], 1-17
System Configuration [With Autoloader], 1-19
Basic Operating Techniques [With Autoloader], 1-21
Carousels, 1-21
Barcode Labels, 1-22
Putting a Barcode Label on a Sample Tube, 1-22
ix
Contents
Putting Sample Tubes in a Carousel, 1-23

Installing a Carousel, 1-24
Removing a Carousel, 1-26
Installing the Plate Adapter, 1-27
Removing the Plate Adapter, 1-27
Consumables and Supplies, 1-27
Reagents, 1-27
Ordering Information, 1-27
Instrument Specifications, 1-28
Dimensions, 1-28
Installation Category, 1-28
Pollution Degree, 1-28
Acoustic Noise Level, 1-28
Electrical Ratings, 1-28
Cytometer, 1-29
Performance Characteristics, 1-31
Reagent Limitations, 1-31
Material Safety Data Sheets (SDS/MSDS), 1-31
CHAPTER 2: Using the CytExpert for DxFLEX Software, 2-1

Overview, 2-1
Launching the Software, 2-1
Main Software Screen, 2-1
Start Page, 2-2
Acquisition Screen, 2-3
Acquisition Screen Navigation, 2-4
Collection, 2-5
Test Tubes, 2-6
Plot area, 2-8
Status Bar, 2-9
Analysis Screen, 2-10
Compensation Experiment Screen, 2-11
QC Experiment Screen, 2-11
QC Report Screen, 2-11
QC Experiment Screen, 2-13
QC Screen Navigation, 2-13
Software Menu, 2-14
Acquisition and Analysis Screen Menu, 2-14
User Management, 2-18
Creating, Deleting, and Modifying Users in User Manager, 2-20
Creating a New User in User Manager, 2-20
Deleting Users in User Manager, 2-21
Modifying Users in User Manager, 2-21
Unlocking a User Account, 2-22
Resetting a User Passwords, 2-22
x
Contents
Changing a User Password, 2-22

Role Management, 2-23
Creating, Deleting, and Modifying User Roles in Role Manager, 2-24
Creating New User Roles in Role Manager, 2-24
Deleting User Roles in Role Manager, 2-25
Modifying User Roles in the Role Window, 2-25
Account Policies, 2-26
User Management Operation Log, 2-29
Viewing and Exporting User Logs, 2-29
Maintenance Log, 2-29
Adding/Deleting a Maintenance Entry, 2-30
Adding/Deleting Maintenance Tasks, 2-32
Adding a Maintenance Task, 2-32
Deleting a Maintenance Task, 2-35
Viewing and Exporting Maintenance Logs, 2-37
Graphic and Gating Styles, 2-39
Plots, 2-39
Gates, 2-40
Software Settings, 2-41
Language Settings, 2-47
CHAPTER 3: Operation Principles, 3-1

Overview, 3-1
Sample Flow, 3-1
Sample Loading, 3-1
Hydrodynamic Focusing, 3-2
Laser Beam Shaping, 3-3
Cell Illumination, 3-4
Forward Scatter, 3-4
Side Scatter and Fluorescent Light, 3-4
Light Collection, Separation and Measurement, 3-4
Forward Scatter Collection, 3-4
Side Scatter and Fluorescent Light Collection, 3-5
Side Scatter, 3-5
Fluorescent Light, 3-5
Signal Processing, 3-6
Data Storage, 3-6
Automated Software Features, 3-6
Parameters, 3-7
TIME Parameter, 3-7
Plot Display, 3-7
Statistics, 3-8
xi
Contents
CHAPTER 4: Daily Startup, 4-1

Overview, 4-1
Pre-Startup Inspection, 4-1
Check Waste and Reagent Levels, 4-2
Power Source Inspection, 4-4
Workstation Connections Inspection, 4-4
Turning On the Instrument, 4-4
Opening the Software, 4-4
Logging Out of the Software, 4-8
Selecting the Proper Sample Injection Mode [Without
Autoloader], 4-8
Using Semi-Automatic Injection Mode, 4-9
Using the Manual Injection Mode, 4-10
Selecting the Proper Sample Injection Mode [with
Autoloader], 4-14
Using Single Tube Mode or Carousel Mode, 4-15
Running the System Startup Program [Without Autoloader], 4-17
Running the System Startup Program [With Autoloader], 4-21
Selecting Experiments from the Start Page, 4-26
Initializing the Instrument, 4-27
CHAPTER 5: Instrument Quality Control and Standardization, 5-1

Overview, 5-1
Preparing the QC Sample Tube, 5-3
Required materials, 5-3
Preparation process, 5-3
Preparing the QC Well Plate, 5-3
Importing Lot-Specific Target Values, 5-4
Collecting QC Data, 5-8
Confirming Results, 5-12
Creating Levey-Jennings Charts, 5-15
QC Result Manager, 5-19
Standardization, 5-20
Generating Target Median Values, 5-21
Managing Standardization Target Library, 5-26
Adding a New Standardization Item, 5-27
Editing Standardization Item Parameters, 5-28
Importing a Standardization Item, 5-29
Deleting Standardization Items, 5-29
Performing the Standardization, 5-29
Applying the Standardized Acquisition Settings, 5-32
xii
Contents
CHAPTER 6: Data Acquisition and Sample Analysis, 6-1

Overview, 6-1
Creating Experiments, 6-1
Creating an Experiment, 6-2
Setting Tube Property [Single Tube or Carousel Mode], 6-3
Worksheet Modes, 6-14
Switching to Shared Worksheets, 6-15
Switch to Independent Worksheets, 6-15
Changing the Tube Name, 6-16
Setting the Channel and Label, 6-16
Creating Plots and Gates, 6-19
Creating and Adjusting Auto Gates, 6-30
Turning Auto Recalculate On/Off, 6-31
Adjusting Autogate Movement and Extent, 6-32
Setting Customized Parameters, 6-34
Setting Custom Statistics, 6-36
Managing the Panel Library, 6-38
Creating a New Panel Experiment, 6-39
Saving a Panel to the Panel Library, 6-39
Importing a Panel, 6-40
Exporting a Panel, 6-41
Deleting a Panel, 6-42
Using Heat Maps, 6-42
Load Sample and Record Data, 6-43
Before Running Samples, 6-43
Verifying, Selecting, Editing, and Creating Detector
Configuration, 6-44
Sampling and Collecting Data, 6-47
Configuring Acquisition Settings, 6-53
Laser Settings, 6-53
Adjusting the Gain, 6-54
Adjusting the Threshold, 6-55
Setting Collection Conditions, 6-57
Setting Plot Display Conditions, 6-59
Analyzing and Exporting Data, 6-60
Exporting FCS Files, 6-66
Exporting Single Tube Files, 6-66
Exporting Multiple FCS Files, 6-68
Exporting Plots or the Statistics Table of Multiple Tubes as Picture
Files, 6-69
Batch Exporting Reports, 6-69
Automatically Exporting Reports, 6-71
Importing and Exporting Instrument Settings, 6-73
Importing Instrument Settings, 6-73
Exporting Instrument Settings, 6-74
Importing and Exporting Compensation Settings, 6-74
Printing Graphics, 6-75
xiii
Contents
Printing Batch Reports, 6-77

Saving the Experiment, 6-78
Concluding the Experiment, 6-78
CHAPTER 7: Compensation, 7-1

Overview, 7-1
Creating a Compensation Experiment, 7-3
Preparing the Compensation Sample, 7-5
Using Control Samples to Generate the Compensation Matrix, 7-6
Defining the Negative Population Using Unstained Samples, 7-6
Running the Single Positive Control Samples, 7-7
Calculating Compensation Values, 7-9
Creating the Compensation Matrix from Previously Acquired Data, 7-11
Adjusting Compensation, 7-12
Manually Adjusting Compensation, 7-12
Importing and Exporting Compensation, 7-13
Importing Compensation Settings from Compensation Matrix
Files, 7-13
Importing Compensation Settings from the Compensation
Library, 7-14
Exporting Compensation Settings, 7-15
Managing the Compensation Library, 7-17
Adding Channels for Compensation, 7-18
CHAPTER 8: Data Review, 8-1

Overview, 8-1
Copying Experiments and Importing Data, 8-1
Copying a Previously Acquired Experiment, 8-1
Importing Previously Acquired Data, 8-1
Setting the Plots and Statistics, 8-3
Opening the Analysis Screen, 8-3
Creating Multi-Data Histograms and Dot Plot Overlays, 8-5
Calculating Sample Injection Volume and Concentration, 8-7
Adjusting Compensation Settings, 8-8
Reports, 8-9
Creating a Report, 8-9
Copying, Deleting, or Renaming a Report, 8-9
Setting a Report as Default, 8-9
Editing the Report Master Page, 8-9
Designing a Report, 8-10
Histogram Overlay, 8-11
Dot Plot Overlay, 8-11
Test Result, 8-12
Managing the Statistics Settings Displayed in the Test Result
xiv
Contents
Element, 8-13
Exporting the Test Result Element as a CSV File, 8-15
Text, 8-16
Image, 8-16
Property, 8-17
Page Number, 8-18
Adding and Deleting Pages from a Report, 8-19
Zooming In and Out of a Report, 8-19
Linking a Plot from a Worksheet to a Report, 8-19
Report Output, 8-21
Analyzing Samples and Tubes, 8-21
Analyzing a Single Sample or Tube, 8-22
Cancelling an Analysis, 8-23
Batch Analyze, 8-24
Canceling Batch Analyses, 8-26
Reviewing Samples and Tubes, 8-27
Reviewing a Single Sample or Tube, 8-28
Canceling a Review, 8-30
Batch Review, 8-30
Canceling Batch Reviews, 8-33
Exporting Results, 8-34
CHAPTER 9: Daily Shutdown, 9-1

Overview, 9-1
Preparing the Cleaning Solution, 9-1
Shutting Down the Instrument, 9-1
CHAPTER 10: Troubleshooting, 10-1

Overview, 10-1
Precautions/Hazards, 10-1
Laser Related Hazards, 10-1
Laser Beam Hazards, 10-2
Laser Warning Labels, 10-2
Hazard Labels and Locations, 10-6
Biohazard Label and Location, 10-6
Electrical Shock Hazard Label and Location, 10-8
Caution Labels and Location, 10-9
Disposal of Electrical Instrumentation, 10-10
RoHS Notice, 10-11
RoHS Caution Label, 10-11
Disposal Precaution, 10-11
Troubleshooting Table, 10-12
Backup/Restore, 10-23
xv
Contents
Backup, 10-23
Restore, 10-25
Viewing Cytometer Information, 10-27
CHAPTER 11: Cleaning Procedures, 11-1

Overview, 11-1
Routine Cleaning, 11-1
Daily Clean, 11-1
Daily Clean [Without Autoloader], 11-2
Daily Clean [With Autoloader - Single Tube and Carousel
Mode], 11-4
Cleaning the Sample Station or Autoloader, 11-7
Cleaning the Sample Probe, 11-8
Deep Clean Procedure, 11-9
Cleaning the Sheath Fluid Container, 11-11
Cleaning the Waste Container, 11-12
Nonscheduled Cleaning, 11-14
Preparing the Instrument for Transport or Storage, 11-14
Lifting and Carrying Instructions [Without Autoloader
Only], 11-15
CHAPTER 12: Replacement/Adjustment Procedures, 12-1

Overview, 12-1
Routine Replacement/Adjustment, 12-2
Front Cover Removal, 12-2
Removal, 12-2
Reinstallation, 12-3
Right-Side Cover Removal and Reinstallation, 12-4
Removal, 12-4
Top and Front Cover of DxFLEX Autoloader Removal and
Removal, 12-7
Filling the Sheath Fluid Container, 12-9
Emptying the Waste Container, 12-10
Managing the Maintenance Reminder, 12-11
Adding the Deep Clean Solution, 12-14
Replacing the Sheath Fluid Filter, 12-16
Replacing the Sample Probe and/or the Sample Peristaltic Pump
Tubing, 12-20
[DxFLEX without Autoloader], 12-20
[DxFLEX with Autoloader], 12-25
Inspecting the Liquid Flow Path for Leaks, 12-33
Priming the Flow Cell, 12-34
Changing the Event Rate Setting, 12-35
xvi
Contents
Nonscheduled Replacement/Adjustment, 12-35

Calibrating the Sample Flow Rate, 12-35
Setting Laser Delay, 12-40
Replacing the Optical Filter, 12-41
Replacing the Fuse, 12-43
Replacing the Sheath Fluid Harness and/or Waste Harness, 12-45
Changing Sample Mixing and Backflush Settings, 12-48
Calibrating the Carrier [DxFLEX With Autoloader], 12-50
Calibrating the Carrier in Single Tube or Carousel Mode, 12-50
APPENDIX A: Instrument Installation, A-1

Overview, A-1
Instrument Transportation and Storage, A-1
Installation Environment Validation, A-1
Worktable, A-2
Ventilation and Cleaning, A-2
Power Source, A-3
Temperature and Humidity, A-3
Waste Disposal, A-4
Unpacking the Instrument and Inspecting the Materials for Defects or
Omissions, A-4
Installing the Instrument and Connecting the Equipment [DxFLEX
Without Autoloader], A-5
Updating and Reinstalling the Software, A-5
Required materials, A-5
Uninstalling the Software, A-5
Updating and Reinstalling the CytExpert for DxFLEX Software, A-6
Starting the Software, A-11
APPENDIX B: Barcode Specifications for the DxFLEX [With Autoloader], B-1

Barcode Sample Identification, B-1
Correct Placement of The Barcode Label, B-1
Barcode Label Specifications, B-2
Label Size and Thickness, B-3
Symbol Dimensions, B-3
Label and Print Quality, B-3
Barcode Error Rate, B-4
Barcode Symbologies, B-4
Barcode Labels, B-5
Barcode Label Optical Characteristics at 650 nm ±10%, B-6
Autoloader Barcode Reader, B-6
Checksum Algorithm, B-7
xvii
Contents
APPENDIX C: DxFLEX [With Autoloader -Plate Mode], C-1

Overview, C-1
Installing the Plate Adapter, C-1
Removing the Plate Adapter, C-4
Using the CytExpert for DxFLEX Software, C-6
Collection, C-6
Test Tubes, C-7
Software Settings, C-9
Selecting the Plate Sample Injection Mode [With Autoloader - Plate
Mode], C-10
Using Plate Mode, C-10
Selecting the Proper Sample Injection Mode [with
Autoloader], C-13
Running the System Startup Program [in Plate Mode], C-13
Instrument Quality Control and Standardization, C-18
Preparing the QC Well Plate, C-18
Required Materials, C-18
Preparation process, C-19
Setting up the Plate, C-19
Collecting QC Data, C-21
Data Acquisition and Sample Analysis, C-21
Setting Tube Property [With Autoloader-Plate Mode], C-21
Sampling and Collecting Data, C-32
Using Heat Maps, C-33
Creating a Heat Map, C-33
Refreshing a Heat Map, C-39
Modifying Existing Heat Map Settings, C-40
Deleting an Existing Heat Map, C-41
Exporting a Heat Map, C-41
Daily Clean [With Autoloader - Plate Mode], C-41
Calibrating the Sample Flow Rate in Plate Mode, C-44
Calibrating the Carrier in Plate Mode, C-48
APPENDIX D: Good Practices for Cyber Security, D-1

Overview, D-1
Drive Encryption, D-1
BitLocker Keys, D-2
Changing Encryption Strength Level for BitLocker, D-2
Changing the encryption strength for BitLocker, D-2
Enabling BitLocker, D-6
Backing Up Your Recovery Key, D-13
BitLocker Decryption, D-17
Turning off BitLocker, D-17
xviii
Contents
Suspending BitLocker Drive Encryption, D-21

Suspending BitLocker , D-21
Restarting BitLocker Protection, D-23
Recovering Your BitLocker Key, D-23
BitLocker Recovery, D-24
Protection from Malware Software, D-25
Installing McAfee Application Control, D-26
Enabling McAfee Application Control, D-30
Exporting the Certificate, D-34
Importing BEC Certificate, D-44
Switching McAfee Application Control to Update Mode, D-46
Disabling McAfee Application Control, D-48
Operating System Updates, D-52
Enabling System Protection, D-52
Creating a Restore Point, D-55
Downloading Operating System Updates, D-56
Notification Options, D-57
Installing the Operating System Updates , D-58
Recovering from Failed Operating System Updates, D-60
System Hardening, D-65
Enabling Automatic Log-Off, D-65
Enabling Screen Saver Protection, D-65
Setting Lock Screen , D-73
Setting Account Lockout Policy, D-75
Setting Password Policy, D-80
Disabling Remote Access, D-83
Disabling Auto Play, D-85
Disabling Unnecessary Services, D-87
Disabling Unauthorized Applications, D-91
Enabling Installation Restriction, D-95
Enabling Firewall Defender, D-97
Enabling Network Time Protocol, D-99
Remote Access, D-103
APPENDIX E: Table of Hazardous Substances, E-1

Table of Hazardous Substances, E-1
Abbreviations
References
Index
Beckman Coulter, Inc.
Customer End User License Agreement
Related Documents
xix
Illustrations
Illustrations
1.1 Main Components [Without Autoloader], 1-2

1.2 Main Components [With Autoloader], 1-3
1.3 Optical Filter Mounts, 1-5
1.4 Optical Filter Mount with Optical Filter, 1-5
1.5 Optical Filter Mount Labeled with the Band-Pass
Information, 1-5
1.6 Optical Filter Mount (Top), 1-6
1.7 Fluid Containers [Without Autoloader Shown], 1-9
1.8 Fluidics Module View, 1-10
1.9 Fluidic Connections, 1-11
1.10 Sample Station, 1-12
1.11 Sample Tube Holder Positions, 1-13
1.12 Sample Station [With Autoloader - Carousel Installed], 1-14
1.13 Sample Station [With Autoloader - Plate Adapter Installed], 1-15
1.14 Autoloader Carousel Barcode Labels, 1-16
1.15 System Connections [Without Autoloader], 1-17
1.16 Back Cover Connections [Without Autoloader], 1-18
1.17 Front of Cytometer [Without Autoloader], 1-18
1.18 System Connections [With Autoloader], 1-19
1.19 Back Cover Connections [With Autoloader], 1-20
1.20 Front of Cytometer [With Autoloader], 1-21
2.1 Drawing Controls Toolbar (Top of Screen), 2-10
2.2 QC Report Screen, 2-12
2.3 Software Menu Tree†, 2-14
2.4 QC Software Menu Tree, 2-14
2.5 User Manager (Card View), 2-18
2.6 User Manager (Grid View), 2-19
2.7 Role Manager, 2-23
2.8 Account Policies - Password Policy, 2-27
2.9 Account Policies - Account Lockout Policy, 2-28
2.10 Account Policies - Application Inactivity Policy, 2-28
3.1 Flow Cell, 3-3
3.2 Laser Beam Shaping, 3-4
3.3 Light Path through the WDM with a Single Port, 3-5
xx
Illustrations
3.4 Time vs Fluorescence plot, 3-7

6.1 Independent Worksheets, 6-15
6.2 Shared Worksheet, 6-15
6.3 All Gates - Example Experiment, 6-27
6.4 Circular Gating Logic - Example Experiment, 6-27
6.5 Movement - Default Setting, 6-33
6.6 Movement - Max Setting, 6-33
6.7 Extent - Default Setting, 6-34
6.8 Extent - Maximum Setting, 6-34
6.9 Panel Library Window, 6-39
6.10 Laser Setting Window, 6-53
7.1 Before Compensation, 7-2
7.2 After Compensation, 7-2
7.3 Positive Population Selected from the Single-Stained
Sample, 7-8
7.4 Positive and Negative Populations Without an Unstained
Sample, 7-9
8.1 Report Toolbar - Standard Experiment, 8-10
8.2 Report Toolbar - Panel Experiment, 8-10
8.3 Test Result Element (Right-Click Menu Shown), 8-12
8.4 Test Result Property Window, 8-12
8.5 Output Options on Report Toolbar, 8-21
10.1 Laser Warning Label on the Laser Optical Bench [DxFLEX
Without Autoloader], 10-3
10.2 Laser Warning Label on the Laser Optical Bench [DxFLEX With
Autoloader], 10-3
10.3 Laser Warning Label within the Optical Bench (Located Inside
the Cytometer) [DxFLEX Without Autoloader], 10-4
10.4 Laser Warning Label within the Optical Bench (Located Inside
the Cytometer) [DxFLEX With Autoloader], 10-4
10.5 Laser Label in Autoloader [DxFLEX With Autoloader], 10-5
10.6 Laser Warning Labels on the Cytometer Back Cover [DxFLEX
10.7 Laser Warning Labels on the Cytometer Back Cover [DxFLEX
With Autoloader], 10-6
10.8 Biohazard Label on the Fluid Containers [DxFLEX Without
Autoloader Shown], 10-6
10.9 Biohazard Label located in the Sample Station and on the Back
of the Cytometer [DxFLEX Without Autoloader], 10-7
10.10 Hazard Label located in the Semi-automatic Sampling Station of
the Cytometer [DxFLEX Without Autoloader], 10-7
xxi
Illustrations
10.11 Biohazard Label located in the Sample Station and on the back
of the Cytometer [DxFLEX With Autoloader], 10-8
10.12 Electrical Shock Hazard Label by the Power Switch [DxFLEX
10.13 Electrical Shock Hazard Label by the Power Switch [DxFLEX
With Autoloader], 10-9
10.14 Caution Labels [DxFLEX Without Autoloader], 10-9
10.15 Caution Labels [DxFLEX With Autoloader], 10-9
10.16 Hazard Labels in the Autoloader, 10-10
10.17 Cytometer Information Window, 10-27
B.1 Barcode Label, B-1
B.2 Barcode Label Placement, B-2
B.3 Barcode Label Specifications, B-3
D.1 gpedit Search, D-2
D.2 Choosing Drive Encryption Method and Cipher Strength, D-4
D.3 Choosing the Encryption Level, D-5
D.4 Checking PC Configurations, D-8
D.5 BitLocker Encrypting the Drive, D-12
D.6 BitLocker Successfully On, D-12
D.7 Security Update Setup Installing, D-59
D.8 Security Update Setup Successfully Installed, D-59
D.9 Windows Shutdown, D-59
D.10 Working on Completing Security Updates, D-60
xxii
Tables
Tables
1.1 WDM Optical Filter Mount Color Codes, 1-7

10.1 Troubleshooting, 10-12
B.1 Barcode Symbologies, B-5
B.2 Code-Related Specifications, B-6
D.1 List of Unnecessary Services, D-87
E.1 Table of Hazardous Substances Name and Concentration, E-2
xxiii
Tables
xxiv
Introduction
Overview
This introduction contains the following information:
• How to Use Your Manual

• About this Manual
• Conventions Used
• Graphics
How to Use Your Manual
Use this Instructions for Use manual for information on the day-to-day operation of your DxFLEX
flow cytometer. You can find detailed step-by-step procedures for Daily Startup and Quality
Control, configuring settings, running samples, analyzing data, and performing Startup and
Shutdown. This manual also contains physical and system specifications, safety and
troubleshooting information, as well as information about what your DxFLEX flow cytometer does
and the methods it uses. It also contains procedures for cleaning and replacement.
About this Manual
The information in your Instructions for Use manual is organized as follows:
CHAPTER 1, System Overview

Provides information regarding the individual components of the DxFLEX flow cytometer and the
corresponding functions of these components.
CHAPTER 2, Using the CytExpert for DxFLEX Software

Provides an overview of each aspect of the software’s functions.
CHAPTER 3, Operation Principles

Describes how the Cytometer measures scattered light and fluorescence as cells pass through the
laser beam.
CHAPTER 4, Daily Startup

Provides instructions for starting your DxFLEX flow cytometer and navigating to the sample testing
standby state.
C44966AB xxv
Introduction
About this Manual
CHAPTER 5, Instrument Quality Control and Standardization

Provides instructions for performing daily quality control (QC) on your DxFLEX flow cytometer to
confirm the instrument is working correctly and to ensure accurate data. Quality control allows you
to determine whether your instrument can provide adequate signal strength and precision.
CHAPTER 6, Data Acquisition and Sample Analysis

Provides instructions for operating the DxFLEX instrument, including data acquisition, analyzing
and exporting results, and manually adjusting the compensation during the acquisition and
analysis.
CHAPTER 7, Compensation
Describes how to create a compensation experiment and automatically calculate compensation
values after acquiring the data. It also explains how to use these calculations for other experiments.
CHAPTER 8, Data Review

Describes how to use the Analysis screen to analyze data that has already been acquired through
the instrument.
CHAPTER 9, Daily Shutdown

Describes how to keep the instrument in optimal condition through daily cleaning during the
shutdown procedure.
CHAPTER 10, Troubleshooting

Describes some common problems and their solutions in a basic troubleshooting matrix.
CHAPTER 11, Cleaning Procedures

Describes how to carry out certain routine and nonscheduled cleaning procedures.
CHAPTER 12, Replacement/Adjustment Procedures

Describes how to carry out certain routine and nonscheduled replacement and adjustment
procedures.
APPENDIX A, Instrument Installation

Provides the instrument setup procedures for your DxFLEX flow cytometer.
APPENDIX B, Barcode Specifications for the DxFLEX [With Autoloader]

Provides the barcode specifications for the DxFLEX [With Autoloader].
APPENDIX C, DxFLEX [With Autoloader -Plate Mode]

Provides the procedures for using the plate adapter and plate mode.
APPENDIX D, Good Practices for Cyber Security

Provides information on cyber security.
APPENDIX E, Table of Hazardous Substances

Provides the table of hazardous substances with the hazardous substance name and concentration.
xxvi C44966AB
Introduction
Conventions Used
Conventions Used
This document uses the following conventions:
• Bold face font indicates buttons or selections that appear on the workstation screen.
• The term “select” is used to indicate the following action:
— To click with a mouse.
NOTE The verb “press” is reserved for mechanical buttons, such as keys on the keyboard.
• The software path to a specific function or screen appears with the greater than (>) symbol
between screen options.
• Links to information in another part of the document for additional information are in blue . To
access the linked information, select the blue, underlined text.
• The information in your Instructions for Use manual applies to both DxFLEX instruments
equipped with an Autoloader and DxFLEX instruments not equipped with an Autoloader unless
otherwise specified.
• Sections that contain entirely new content are flagged with a New Section icon at the end
of the section title.
NOTE When information in this document only applies to the Autoloader configuration, it is marked
[With Autoloader]. When information in this document only applies to the instrument without an
Autoloader, it is marked [Without Autoloader].
IMPORTANT IMPORTANT is used for comments that add value to the step or procedure being performed.
Following the advice in the IMPORTANT adds benefit to the performance of a piece of equipment or to a
process.
NOTE NOTE is used to call attention to notable information that should be followed during use, or
maintenance of this equipment.
Graphics
All graphics, including screens and printouts, are for illustration purposes only and must not be
used for any other purpose. For example, software screens that show the DxFLEX system in the
background may not depict the latest production version of the system.
C44966AB xxvii
Introduction
Graphics
xxviii C44966AB
CHAPTER 1
System Overview
Overview
This chapter describes the individual components of the DxFLEX flow cytometer and the
corresponding functions of these components.
This chapter contains information on:
• Intended Use
• Main Components
• Optical Components
• Fluidics System
• Sample Station [Without Autoloader]
• Sample Station [With Autoloader]
• System Configuration [Without Autoloader]
• System Configuration [With Autoloader]
• Basic Operating Techniques [With Autoloader]
• Consumables and Supplies
• Instrument Specifications
• Performance Characteristics
• Reagent Limitations
Intended Use
The DxFLEX flow cytometer provides a quantitative and multi-parametric analysis quickly on
suspended cells or other bio-particles at a cell level. For in vitro diagnostic use.
C44966AB 1-1
System Overview
Main Components
Main Components
CAUTION
Risk of instrument damage and/or instrument instability. Do not place any objects
on top of the instrument, as this could cause warping of the top cover or affect the
stability of the optical path.
The instrument consists of Fluid Containers and a Cytometer (including CytExpert for DxFLEX
software). The Autoloader and Plate Adapter for Autoloader are optional accessories.
Figure 1.1 Main Components [Without Autoloader]
b c d
1. Fluid Containers. Accommodates sheath fluid and waste liquids as required for
operation of the instrument.
2. Cytometer (including CytExpert for DxFLEX software). Generates and collects
signals.
3. Workstation. Displays and controls workstation content, and displays data generated
by the Cytometer.
1-2 C44966AB
System Overview
Optical Components 1
Figure 1.2 Main Components [With Autoloader]
upper upper
lower
b c d e
1. Cytometer (including CytExpert for DxFLEX software). Generates and collects
signals.
2. Autoloader. Sample processing area. The Autoloader supports either a 32-tube
carousel or a plate adapter to hold a 96-well plate.
NOTE The Autoloader is an optional accessory that can be purchased separately.

NOTE The plate adapter is an optional accessory that can be purchased separately.
NOTE The plate adapter is not validated for use the DuraClone B27 Reagent Kit.
3. Fluid Containers. Accommodates sheath fluid and waste liquids as required for
operation of the instrument.
4. Workstation. Displays and controls workstation content, and displays data generated
by the Cytometer.
Optical Components
CAUTION
Risk of operator injury. When operating the instrument, keep the top cover in the
closed position to prevent the top cover from falling. When opening the top cover,
be cautious to avoid any possible pinch points.
The optical components are located in the upper portion of the Cytometer and are visible when the
top cover is open. Three parts are included: an optical bench, detector arrays also known as
wavelength division multiplexers (WDMs), and optical fibers. Optical components include
C44966AB 1-3
System Overview
Optical Components
equipment such as lasers and signal detectors that are used to excite, transmit, and collect optical
signals.
Optical Components [Without Autoloader Shown]
d
c
b
e
CAUTION
CLASS 3B LASER RADIATION WHEN
OPEN AND INTERLOCKS DEFEATED
AVOID EXPOSURE TO BEAM
1. Optical bench. Includes laser light sources, an optical beam combiner, and an integrated optics flow
cell assembly. The optical bench cover is equipped with a laser interlock that turns the lasers off unless
the cover is tightly closed.
2. Wavelength division multiplexer (WDM). Each WDM is a unique detector array that corresponds to a
different laser. Each WDM contains optical filters and detectors for detecting channel fluorescence or
scatter from a particular laser. It is necessary to ensure that the filter and software settings match for
each channel.
3. Optical fiber. Transmits emitted fluorescence specific to laser path.
CAUTION
Risk of instrument damage. Do not place sample tubes in the optical filter holder. Liquid spills can
damage instrument components. Use a tube rack to hold any sample tubes.
4. Optical filter holder. Securely holds additional DxFLEX optical filters.
Wavelength Division Multiplexer (WDM)

Each WDM corresponds to a different laser. The color of the ring on each cap corresponds to the
color of the respective laser. Pressing the two release buttons on opposite edges of the cap allows
you to open the unit and replace the filters inside. See Figure 1.3. All optical filters are designed to
1-4 C44966AB
System Overview
be interchangeable. Refer to Replacing the Optical Filter in CHAPTER 12, Replacement/Adjustment

Procedures, to replace an optical filter.
Figure 1.3 Optical Filter Mounts
Each optical filter mount has an optical filter glass piece. See Figure 1.4.
Figure 1.4 Optical Filter Mount with Optical Filter
1. Optical filter glass piece
Each optical filter mount is labeled with the corresponding laser and band-pass information. See
Figure 1.5.
Figure 1.5 Optical Filter Mount Labeled with the Band-Pass Information
C44966AB 1-5
System Overview
Optical Components
The top of each optical filter mount has two marks. The color of the dot (1) indicates the color of
the laser. See Figure 1.6. The color of the line (2) indicates the wavelength range of the optical
band-pass filter. See Figure 1.6.
Figure 1.6 Optical Filter Mount (Top)
1. . Indicates corresponding laser color: Blue indicates a 488 nm laser; Red indicates a
638 nm laser; Violet indicates a 405 nm laser.
2. . Indicates the band-pass wavelength ranges; the specific wavelength is

indicated numerically on the lateral side of the mount.
Band-pass filters are used to transmit fluorescence at the specific wavelength ranges. These ranges
are designed to measure fluorescence from the fluorochromes such as those listed in Table 1.1.
(with red and violet laser upgrades installed) that are excited by the lasers. You can change the
optical filters according to your detector configuration. There is no need to realign the optical
system when the filters are changed.
1-6 C44966AB
System Overview
Table 1.1 WDM Optical Filter Mount Color Codes
DxFLEX
Channel Commonly used Fluorescent
Laser Fluorescent Channel Names Dyes
FITC FITC, Alexa Fluor™ 488, CFSE,

488-nm 525/40 BP Fluo-3
PE PE, PI
585/42 BP
ECD ECD, PE-Texas Red®,
610/20 BP PE-CF594, PI
PC5.5 PC5.5, PC5, PerCP,
690/50 BP PerCP-Cy5.5, PI
PC7 PC7
780/60 BP
APC APC, Alexa Fluor™ 647,
638-nm 660/10 BP eFluor™ 660
APC-A700 APC-A700, Alexa Fluor™700
712/25 BP
APC-A750 APC-A750, APC-Cy7, APC-H7,
780/60 BP APC- eFluor™ 780
PB450 Pacific Blue™dye, V450,
405-nm 450/45 BP eFluor™ 450, BV421
KO525 Krome Orange, AmCyan, V500,
525/40 BP BV510
Violet610 BV605, Qdot® 605
610/20 BP
660/10 BP
780/60 BP
NOTE The FITC channel, PE channel, and PC5.5 channel are validated for IVD use with the DuraClone B27
Reagent.
Optical Fiber
CAUTION
Risk of data integrity damage.
• During use, verify that the optical fibers are securely connected to the WDM.
A loose connection can alter the optical path and affect fluorescence
detection.
• Do not disconnect the fiber as this could contaminate the tip and weaken the
signal.
Fluorescence emitted by laser-excited fluorochromes is picked up and delivered by each optical

fiber to the corresponding detector module. Each optical fiber has a colored ring on the end that
C44966AB 1-7
System Overview
Fluidics System
connects to the WDM, indicating the color of the corresponding laser. Ensure that the correct fiber
is properly connected to the corresponding WDM.
Optical Fiber [Without Autoloader Shown]

q w e
CAUTION
1. Red laser fiber

2. Violet laser fiber
3. Blue laser fiber
Fluidics System
The fluidics system consists of two parts: the Fluid Containers and the Fluidics module. The Fluidics
module is located on the right side of the Cytometer. You need to open the right-side cover of the
instrument (see Right-Side Cover Removal and Reinstallation in CHAPTER 12,
Replacement/Adjustment Procedures) to perform maintenance operations. The fluidics system
helps to transmit the sheath fluid at a stable rate into the flow cell, forming a laminar fluidics
system that ensures that the tested particles go through the detection area sequentially.
1-8 C44966AB
System Overview
Fluidics System 1
IMPORTANT The fluid containers should be placed on the left side of the cytometer when using a DxFLEX
[Without Autoloader].
The fluid containers should be placed directly behind the Autoloader when using a DxFLEX [With
Autoloader].
Figure 1.7 Fluid Containers [Without Autoloader Shown]
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1. Fluid Container holder. Holds the Fluid Containers.

2. Fluid sensor holder cutout. Holds the sheath fluid harness and the waste harness when removed from
their respective containers to protect the sensors from damage and/or contamination.
3. Sheath fluid harness. Connects to the sheath fluid container; conveys the sheath fluid into the
instrument. The sheath fluid harness includes a level sensor, sheath fluid tubing, and backflush tubing.
The other end of the harness is connected to the Fluidics module in the Cytometer. When the sheath
fluid container is near empty, a warning notice is transmitted to the instrument and an audible signal
sounds as a warning.
4. Sheath fluid container. 4 L capacity, for holding sheath fluid. Beckman Coulter recommends using
DxFLEX Sheath Fluid or a similar nonionic strength sheath fluid to ensure system performance.
5. Waste harness. Connects to the waste container; conveys the waste liquid from the instrument to the
waste container. The waste harness includes a level sensor. The other end of the harness is connected
to the Fluidics module in the Cytometer. When the waste container is near full, a warning notice is
transmitted to the instrument and audible signal sounds as a warning.
6. Waste container. 4 L capacity, for holding waste liquids. Attention to biosafety and waste labeling is
required.
Fluid Containers
Two Fluid Containers are placed in the Fluid Container holder: a sheath fluid container and a waste
container. See Figure 1.7. Each container cap is fitted with a harness and a level sensor. The blue
harness connects to the sheath fluid container, while the yellow harness connects to the waste
container. The containers do not require additional pressure inside. Take all necessary biosafety
precautions and use proper personal protective equipment when using the Fluid Containers.
C44966AB 1-9
System Overview
Fluidics System
Fluidics Module
The Fluidics module is on the right side of the Cytometer. To access it, you must first open the
right-side cover. Refer to Right-Side Cover Removal and Reinstallation in CHAPTER 12,
Replacement/Adjustment Procedures. Inside the module, in addition to working pumps, valves,
and tubing, there is a sheath filter and a Deep Clean solution bottle. During maintenance, it may be
necessary to replace the filter (see Replacing the Sheath Fluid Filter in CHAPTER 12,
Replacement/Adjustment Procedures) or to add Deep Clean solution (see Adding the Deep Clean
Solution in CHAPTER 12, Replacement/Adjustment Procedures).
Figure 1.8 Fluidics Module View
PV5 PV1 PV3 PV2
c d e
1. Alarm. Emits a warning sound when there is a problem with the Fluid Container capacity or with the
performance of certain operations.
NOTE When the alarm sounds, the Mute Alerter icon appears in the status bar. The alarm continues
for about 30 seconds. To mute the alarm temporarily select the Mute Alerter in the status bar.
The icon disappears when the waste container is emptied and/or the sheath container is filled.
2. Deep Clean solution peristaltic pump. Transfers cleaning solution to the flow cell based on flow rate
settings.
3. Deep Clean solution bottle. Contains the diluted cleaning solution that helps to clean the flow cell.
4. Sheath fluid filter. 0.2 μm filter, for filtering sheath fluid.
1-10 C44966AB
System Overview
Fluidics System 1
Figure 1.9 Fluidic Connections
b c d
e f g
1. Waste level sensor connector. Connects to the waste liquid sensor cable.
2. Waste out. Connects to the waste tubing of the wash station.
3. Sheath fluid level sensor connector. Connects to the sheath fluid sensor cable.
4. Waste out. Connects to the waste tubing of the flow cell.
5. Sheath return. Return sheath to the sheath tank to decrease the flow fluctuation.
6. Sheath fluid in. Connects to the sheath fluid tubing to deliver sheath to the flow cell.
NOTE Use DxFLEX Sheath Fluid or other filtered nonionic strength sheath fluid. Using unfiltered sheath fluid
can shorten the service life of the sheath fluid filters and increase noise and debris detection.
C44966AB 1-11
System Overview
Sample Station [Without Autoloader]
Sample Station [Without Autoloader]
WARNING
Risk of biohazardous contamination and/or instrument damage. When running
samples, it is important to insert the sample tube all the way down into the sample
tube holder, until the bottom of the sample tube touches the base of the holder.
Failing to do this could cause the sample probe to bend or break on entry. Sample
tubes must not exceed 80 mm in height and the outside diameter must not exceed
13 mm.
Figure 1.10 Sample Station
1. Sample tube holder. Supports sample tubes for testing, such as 12 x 75 mm, 1.5-mL, and 2-mL
microtubes.
2. Sample probe. Draws and transfers samples into the flow cell.
3. Wash station and mixer. During the sampling process, samples are automatically shaken and mixed
for a default time of 1 second. The sample probe is automatically cleaned when the instrument
performs a backflush.
1-12 C44966AB
System Overview
Sample Station [With Autoloader] 1
Sample Tube Holder Positions [Without Autoloader]

Three of the sample tube holder positions are shown in Figure 1.11: sample loading position (1),
standby position (2), and sample acquisition position (3). You can only distinguish the mixing
position from the sample acquisition position if you are looking directly at the sample tube holder
while the Cytometer is processing a sample. The mixing position is about 6 mm lower than the
sample acquisition position.
Figure 1.11 Sample Tube Holder Positions
b
c d
Sample Station [With Autoloader]
WARNING
Risk of biohazardous contamination and/or instrument damage. When running
samples, it is important to insert the sample tube all the way down into the sample
tube holder, until the bottom of the sample tube touches the base of the holder.
Failing to do this could cause the sample probe to bend or break on entry. Sample
tubes must not exceed 80 mm in height and the outside diameter must not exceed
13 mm.
The Autoloader is an automated sample loader for the instrument. It uses a carousel that holds
thirty-two 12 x 75 mm test tubes. Refer to Figure 1.12. A plate adapter can be purchased separately
that holds one 96-well plate (U-bottom, V-bottom, or Flat bottom). Refer to Figure 1.13.
The Autoloader mixes each sample before analysis. You can use the Autoloader to automatically
analyze multiple samples or analyze single tubes.
C44966AB 1-13
System Overview
Figure 1.14 shows the location of the carousel number, tube position, and sample tube barcode
labels on the Autoloader carousel.
Figure 1.12 Sample Station [With Autoloader - Carousel Installed]
b c
1. Autoloader
2. Carousel
1-14 C44966AB
System Overview
Sample Station [With Autoloader] 1
Figure 1.13 Sample Station [With Autoloader - Plate Adapter Installed]
b c
1. Autoloader
2. Plate Adapter
NOTE For installing or removing the plate adapter, refer to APPENDIX C, DxFLEX [With Autoloader -Plate
Mode].
NOTE The plate adapter is not validated for IVD use with the DuraClone B27 Reagent.
C44966AB 1-15
System Overview
Figure 1.14 Autoloader Carousel Barcode Labels
c d
1. Sample tube barcode label

2. Tube position barcode label
3. Carousel ID barcode label
1-16 C44966AB
System Overview
System Configuration [Without Autoloader] 1
System Configuration [Without Autoloader]
CAUTION
Risk of data loss and/or instrument damage. Never shut off the power or
disconnect a data cable while the Cytometer is in the process of performing a task.
This could cause data loss or damage to the system.
Figure 1.15 System Connections [Without Autoloader]
b f
CLASS 1 LASER PRODUCT

COMPLIES WITH 21 CFR 1040.10 AND 1040.11
EXCEPT FOR DEVIATIONS PURSUANT TO
LASER NOTICE NO. 50 DATED JUNE 24, 2007
MANUFACTURED
REF
SN
EC REP
IVD
c d
e
upper upper
g lower
1. Monitor 4. Computer
2. Mouse 5. Cytometer
3. Keyboard 6. Fluid Container holder
C44966AB 1-17
System Overview
System Configuration [Without Autoloader]
Figure 1.16 Back Cover Connections [Without Autoloader]

MANUFACTURED
b
REF
SN
EC REP
IVD
c
d
1. Power switch. Turns Cytometer on and off. An indicator light glows when the power is on.
2. Fuse. Protects the internal system from damage by high electrical current.
3. Power line socket. Supplies the power to the Cytometer.
Figure 1.17 Front of Cytometer [Without Autoloader]
1. Load button. In addition to the software controls, this button can be used for automatic sample loading
and data recording.
1-18 C44966AB
System Overview
System Configuration [With Autoloader] 1
System Configuration [With Autoloader]
CAUTION
Risk of data loss and/or instrument damage. Never shut off the power or
disconnect a data cable while the Cytometer is in the process of performing a task.
This could cause data loss or damage to the system.
Figure 1.18 System Connections [With Autoloader]
b d
c CLASS 1 LASER PRODUCT

MANUFACTURED
REF
SN
EC REP
IVD
REF
IVD
SN
e
EC REP
f
g
upper upper
h lower
1. Monitor 5. Keyboard
2. Autoloader 6. Computer
3. Cytometer 7. Fluid containers
4. Mouse
C44966AB 1-19
System Overview
System Configuration [With Autoloader]
Figure 1.19 Back Cover Connections [With Autoloader]
b CLASS 1 LASER PRODUCT

MANUFACTURED
c
REF
SN
EC REP
d
IVD
REF
e
IVD
SN
f
EC REP
1. USB Connector for Autoloader. Connects the Autoloader to the Workstation.

2. Power switch. Turns Cytometer on and off. An indicator light glows when the power is on.
3. Fuse. Protects the internal system from damage by high electrical current.
4. Power line socket. Supplies the power to the Cytometer.
5. USB Connector for Cytometer. Connects the Cytometer to the Workstation.
1-20 C44966AB
System Overview
Basic Operating Techniques [With Autoloader] 1
Figure 1.20 Front of Cytometer [With Autoloader]
B
1. Load button. In addition to the software controls, this button can be used for automatic sample loading
and data recording.
NOTE The load button can be used on the DxFLEX [Without Autoloader] regardless of sample
injection mode or on the DxFLEX [With Autoloader] when using the Single Tube sample injection
mode.
2. Autoloader indicator light. Notifies users about the status of the autoloader. A solid green light
indicates the autoloader is ready for use. A flashing green light indicates that the autoloader is in use.
A yellow light indicates that the autoloader cover is open during sample acquisition, daily clean, or when
in standby mode. A red light indicates a critical error that has caused the autoloader to stop running.
NOTE Ensure the Autoloader cover is closed during sample acquisition and daily clean. Opening the
Autoloader cover while the system is running causes the Autoloader to stop moving.
Basic Operating Techniques [With Autoloader]
Carousels
The DxFLEX Flow Cytometry Autoloader starter kit contains two carousels, each with 32 tube
positions.
C44966AB 1-21
System Overview
Barcode Labels
CAUTION
Sample misidentification can occur from the use of incorrect, poor quality,
damaged, dirty or improperly placed barcode labels. Follow the Barcode
Specifications for the DxFLEX [With Autoloader] to create your barcode labels to
prevent incorrect sample identification. Barcode label incorrectly placed on
sample tubes could cause misidentified tubes. To prevent misidentified samples,
affix the barcode label as shown below so the MCL can read the label.
You can put a barcode label on each sample tube. See APPENDIX B, Barcode Specifications for the
DxFLEX [With Autoloader] for additional information.
NOTE Barcode labels are not required on sample tubes for system operation. When using the check tube
barcode function, you must input the barcode information manually or using an external barcode reader.
The check barcode function on the Autoloader only checks that the barcode information matches with
the information that was inputted as the Sample ID in the software.
NOTE Beckman Coulter recommends the use of Polypropylene tubes to ensure accurate barcode reading.
Putting a Barcode Label on a Sample Tube
1 Carefully align the label with the tube.
2 Press the label down securely, including edges and corners, without wrinkles or folds.
1. 20 mm (0.79 in.) minimum

2. <7.5 degrees. >20mm
<7.5°
1-22 C44966AB
System Overview
Putting Sample Tubes in a Carousel
• The orientation of a tube with a

bar-code label (1) does not matter. The
Autoloader rotates the tube to find the
bar-code label.
• Do not skip tube positions within a
b
panel. The DxFLEX flow cytometry
Autoloader does not skip a protocol in a
panel when a carousel tube position is
empty. If you lose a sample, delete that
protocol from the panel using the Delete
Tube icon.
Delete Tube - Standard Experiment
Delete Tube - Panel Experiment
• You can skip tube positions to separate

two panels based on your tube location
setup in the Set Location window. Refer
to Creating Experiments in CHAPTER 6,
Data Acquisition and Sample Analysis.
C44966AB 1-23
System Overview
Installing a Carousel
1 Open the Autoloader cover.
IMPORTANT Ensure the carousel is properly seated.
2 Line up the carousel with its turntable, and then gently push down. The carousel is in home
position when the handle points toward the front.
e
c
1. Carousel
2. Carousel hub
3. Carousel hub tab
4. Carousel slot
1-24 C44966AB
System Overview
IMPORTANT The sample tube must be placed in tube position 24 when using the Single Tube sample
injection mode.
3 Place the sample tubes in the correct positions.
4 Close the Autoloader cover.
C44966AB 1-25
System Overview
Removing a Carousel
2 Remove the carousel.
1-26 C44966AB
System Overview
Consumables and Supplies 1
Installing the Plate Adapter

Refer to Installing the Plate Adapter in APPENDIX C, DxFLEX [With Autoloader -Plate Mode].
Removing the Plate Adapter

Refer to Removing the Plate Adapter in APPENDIX C, DxFLEX [With Autoloader -Plate Mode].
Consumables and Supplies
Reagents
The following reagents are required for the DxFLEX instrument:
DxFLEX Daily QC Fluorospheres

DxFLEX Daily QC Fluorospheres is a suspension of fluorescent microspheres which may be used for
daily verification of the DxFLEX flow cytometer’s optical alignment and fluidics system.
DxFLEX Sheath Fluid

A nonionic, non-fluorescent, and azide-free sheath fluid for use on Beckman Coulter DxFLEX flow
cytometers.
Contrad 70® Reagent

Diluted 1:1 with DI water for use in the Deep Clean solution bottle.
FlowClean
For use as a cleaning agent for flow cytometer components that come in contact with blood
samples.
Ordering Information
Your instrument may be upgraded to a more highly configured model. The cost to upgrade depends
on the type of configuration needed. For information on specific upgrades, replacement parts, or
supplies, visit the Beckman Coulter website at any time: www.beckman.com. Otherwise, contact us.
C44966AB 1-27
System Overview
Instrument Specifications
Dimensions
Dimensions
Cytometer [Without 42.5 cm x 42.5 cm x 34 cm
Autoloader]
Instrument Fluid Containers and Fluid 14 cm x 35.6 cm x 35.6 cm
dimensions (Length x Container holder [Without
Width x Height) Autoloader]
Cytometer [With 72 cm x 43.5 cm x 34 cm
Autoloader]
Cytometer [Without 23.4 kg
Autoloader]
Cytometer Weight Cytometer [With 35.8 kg
Autoloader]
Plate Adapter 1.2 kg
Installation Category
Installation Category 2
Pollution Degree
Pollution Degree 2
Acoustic Noise Level

Semi-automatic injection mode: <53 dBA
Carousel mode: <55 dBA
Plate mode: <55 dBA
Measure level (maximum): < 60 dBA
Electrical Ratings
Voltage: AC 100 V ~ 240 V ± 10%,
Frequency: 50 Hz/60 Hz ± 1 Hz
1-28 C44966AB
System Overview
Instrument Specifications 1
Peak Power: 250 VA
Working Power:
• Semi-automatic injection mode: <115 VA

• Carousel mode: <136 VA
• Plate mode: <140 VA
Cytometer
Optics
Excitation Optics The DxFLEX system can be configured with up to three spatially-separated lasers.
The optical system is alignment free. The laser delays are automatically adjusted by
the daily QC system, if required. No user intervention is required to ensure optimum
system performance.
Emission Optics Patent-pending alignment-free integrated optics quartz flow cell design with
>1.3 NA.
Flow Cell dimensions: 430-μm x 180-μm internal diameter.
Laser devices Standard wavelengths Blue laser
• Wavelength: 488-nm, 50 mW
• Beam spot size: 5 μm x 80 μm
Red laser
• Wavelength: 638-nm, 50 mW
Violet laser
• Wavelength: 405-nm laser device, 80 mW
Forward scatter Si-photodiode with built-in 488/8 band-pass filter.
detection
Fluorescence and Fluorescence and side scatter light collected by the objective lens is delivered by
side scatter fiber optics to a patent-pending design with high performance, solid-state, high
detection efficiency, low-noise detector arrays.
Reflective optics with a single transmission band-pass filter in front of each detector.
Side-scattered light resolution 300 nm.
Fluidics System
Sample loading speed Defaults Slow 10 μL/min
Medium 30 μL/min
Fast 60 μL/min
Fluid capacity Standard 4-L sheath fluid and waste containers
C44966AB 1-29
System Overview
Fluidics System
Programmed Initialize, Standby, system startup program, sample mix, backflush, prime, Daily
maintenance cycles Clean, Deep Clean
Sample input formats DxFLEX [Without 5 mL (12 x 75 mm) polystyrene and polypropylene sample
Autoloader] tubes
Microcentrifuge 1.5 mL and 2 mL sample tubes
DxFLEX [With 5 mL (12 x 75 mm) polystyrene and polypropylene sample
Autoloader] tubes
Autoloader-Plate Adapter: 96-well plate (Flat-Bottom,
V-Bottom, U-Bottom)
Electronics
Signal processing Fully digital system with an active range of 7 orders of magnitude, signal
collection speed of 30,000 particles/second (includes 15 parameters)
Digital sampling rate 25 Mhz
Signal Pulse area, height for every channel, width for one selectable channel
Data Management
Software CytExpert for DxFLEX
Language English and Chinese
FCS format FCS 3.0
Minimum Operating systema Windows® 10 Enterprise 2019 LTSC x64-bit
Workstation/computer
Processor 4th Gen Intel® Core™ i3 (3MB Cache, 3.40 GHz)
requirements
Memory 4 GB RAM
Storage 500 GB
Port 1 GB Ethernet port
USB 4 USB 3.0 ports (compatible with USB 2.0)
Compensation Full matrix compensation, manual and automatic.
Novel Compensation Library for storage of spillover values of dyes to easily
determine the correct compensation matrix with new gain settings.
QC Auto daily QC routine with Levey-Jennings tracking and logging.
a. Only Windows 10 Enterprise (2019 LTSC x64 bit) has been validated to work with the DxFLEX workstation.
1-30 C44966AB
System Overview
Performance Characteristics 1
Performance Characteristics
Performance
Sensitivity MESF FITC: 30 molecules of equivalent soluble
fluorochrome (MESF-FITC)
PE: 10 molecules of equivalent soluble
fluorochrome (MESF-PE)
Fluorescence CV 2%
resolution
Forward and side Scatter performance is optimized for resolving the white blood cell subsets
scatter resolution (lymphocytes, monocytes, and granulocytes), red blood cells, and platelets.
Beads Carryover ≤0.5%
Dye Carryover <10%a
a. Fluorescence carryover on the DxFLEX system was assessed by analyzing the PE channel median fluorescence shift of an
unstained quality control sample (Immuno-Trol cells) after the acquisition of a sample stained with Propidium Iodide dye.
Firstly, acquisition of three tubes of unstained Immuno-Trol cells, then three tubes of Immuno-Trol cells stained with 25 μL of
Propidium Iodide (1mg/mL solution) were acquired, the system was cleaned according to the Routine Cleaning Procedure.
Subsequently, three tubes of unstained Immuno-Trol cells were analyzed finally. The average of PE median fluorescence shift
obtained in the unstained sample acquisitions was less than 10%.
Performance [With Autoloader]

Throughput 23 minutes for 32 tubes with 20 second acquisition, 3 second mixing, and 3
(Carousel) second backflush
Throughput (Plate) 35 minutes for 96-well plate with 10 second acquisition, without mixing and
backflush
42 minutes for 96-well plate with 10 second acquisition, 3 second mixing per 5
minutes, and 3 second backflush
Dead Volume 25 μL
(Carousel)
Dead Volume (Plate) 20 μL
Reagent Limitations
Only use nonionic strength sheath fluid, like DxFLEX Sheath Fluid. Do not use sheath fluid
containing electrolytes.
Material Safety Data Sheets (SDS/MSDS)
To obtain an SDS or MSDS for reagents used on the DxFLEX system:

• On the Internet, go to www.beckman.com:
1. Select Safety Data Sheets (SDS/MSDS) from the Support menu.
C44966AB 1-31
System Overview
Material Safety Data Sheets (SDS/MSDS)
2. Follow the instructions on the screen.

3. Contact us if you have difficulty locating the information.
• If you do not have Internet access contact us.
1-32 C44966AB
CHAPTER 2
Using the CytExpert for DxFLEX Software
Overview
The CytExpert for DxFLEX software is a full-feature software package that controls the instrument's
operation, collection of experimental data, and analysis of the results. This chapter will explain the
software’s functions and features.

• Launching the Software
• Main Software Screen
• User Management
• Role Management
• Account Policies
• User Management Operation Log
• Maintenance Log
• Graphic and Gating Styles
• Software Settings
Launching the Software
Select the CytExpert for DxFLEX desktop icon to launch the CytExpert for DxFLEX software.
If there is no desktop shortcut, run the “CytExpert for DxFLEX.exe” software directly from the
software installation directory. The default installation path is C:/Program Files/CytExpert for
DxFLEX.
Refer to Opening the Software in CHAPTER 4, Daily Startup, for detailed instructions on logging into
the software and confirming the connection status.
Main Software Screen
Hover your cursor over any button to display a text pop-up of the button’s function.
C44966AB 2-1
Start Page
The login window automatically opens after the software has been launched. The start page
automatically opens after logging into the software.
NOTE You must change the default password immediately upon initial login. For password requirements,
refer to Changing a User Password.
The following operations can be selected from the start page:
• New Experiment. For creating a new experiment for single tubes. The process creates a file with
the .xit extension and a folder with the same file name where the raw data (.fcs files) are kept.
• New Panel Experiment. For creating a new experiment containing a list of samples,
sample-specific tubes, any parameter labels defined for each tube, and tube-specific cytometer
settings. A sample is a collection of tests, reagents, or fluorescence commonly used together in
2-2 C44966AB
Main Software Screen 2
an experiment. The process creates a file with the .xitp extension and a folder with the same
file name where the raw data (.fcs files) are kept.
NOTE Panel experiments can also be saved as a template (.xitpm file).
• New Experiment From Template. For creating an experiment using previously selected
elements.
• New Compensation. For setting up compensation for an experiment.
• Open Experiment. For opening a previously created experiment.
• Open Compensation. For opening a previously created compensation experiment.
• Exit. For exiting CytExpert for DxFLEX software.
The Experiment, Template, and Compensation tabs below give you the option of opening one of the
10 most recently opened experiments.
C44966AB 2-3
Acquisition Screen
Selecting New Experiment, New Panel Experiment, New Experiment From Template, or Open
Experiment automatically opens the Acquisition screen. The Acquisition screen can be accessed by
selecting on the left side of the page.
1. Navigation. Gives the option of accessing the acquisition screen or analysis screen.
2. Menu. Allows you to configure settings for sample acquisition, instrument operation, and software
options.
3. Instrument Operation Controls. Controls sample loading/unloading and data acquisition and
recording.
4. Collection. Establishes control over data recording options and displays the acquisition status.
5. Test tubes. Allows you to configure and duplicate sample tubes, set display attributes, manage
experimental data and compensation.
NOTE The Tube section of the screen can be expanded or retracted by dragging the top border of
the Tube section of the screen. Expanding this section covers other elements of the screen,
including: Events to Display, Events/Sec, and the Acquisition buttons.
6. Plot area. Includes plot and gating controls, as well as an area for drawing plots and generating graphs.
7. Status bar. Displays instrument connection status and system information.
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Acquisition Screen Navigation

The Acquisition screens have two navigation icons, one for the Acquisition screen and the other for
the Analysis screen.
1. Acquisition screen icon. Accesses the Acquisition screen.

2. Analysis screen icon. Accesses the Analysis screen.
C44966AB 2-5
Collection
Standby state [Single Tube Mode] Initialized state [Single Tube Mode]
Standby state [Carousel Mode] Initialized state [Carousel Mode]
Refer to Collection in APPENDIX C, DxFLEX [With Autoloader -Plate Mode] for the collection states
in the Plate Mode.
2-6 C44966AB
1. Acquisition control. Controls sample loading/unloading and data acquisition and recording.
2. Acquisition status. Displays such information as the acquisition rate (Events/Sec), cell count, duration,
and abort (%).
3. Acquisition conditions. Sets the necessary conditions for recording data.
4. Sample flow rate. Sets the acquisition rate for data collection.
NOTE High acquisition rate increases the abort rate and measurement CVs.
Explanation of restrictions:
Events displayed: 1-500,000
Events recorded: 1-100,000,000
Recording time: ≤1800 seconds
C44966AB 2-7
Test Tubes
[Carousel Mode or Single Tube Mode]
Refer to Test Tubes in APPENDIX C, DxFLEX [With Autoloader -Plate Mode] for the Plate Mode Test
Tubes section description.
2-8 C44966AB
1. Test tube management controls. Manages sample tubes. Used to add, copy, or delete attributes, open the tube
property and open the compensation matrix.
2. Test tube status indication. Displays a colored symbol in front of each tube indicating the status of the tube
processing.
• indicates that the tube data was not collected.
• indicates that the tube data was acquired by selecting Run but not saved, and the tube data can be
overwritten.
• indicates that tube data was reacquired by selecting Run.
• indicates that the tube data was recorded, and this data cannot be overwritten.
• indicates that tube data was reacquired and recorded.
• indicates imported FCS data.
NOTE to the left of the test tube status indication symbol indicates that the sample has been
compensated.
• indicates the data file is missing or there is an error in the data file.
• indicates the Sample ID information does not match with the recorded tube barcode.
• indicates the data has been reviewed.
• indicates that tube data was not collected but was linked to Acq. Settings, and the compensation value
changes with the gain setting.
• indicates that tube data was not collected but was linked to Acq. Settings and was reviewed. The
compensation value changes with the gain setting
• indicates that tube data was not collected but was linked to Acq. Settings, and the compensation value
does not change with the gain setting.
• indicates that tube data was not collected but was linked to Acq. Settings and was reviewed. The
compensation value does not change with the gain setting.
• indicates that tube data was acquired by selecting Run and was analyzed.
• indicates that tube data was acquired by selecting Run and was reviewed.
• indicates that tube data was acquired by selecting Record or Auto Record, and was analyzed.
• indicates that tube data was acquired by selecting Record or Auto Record, and was reviewed.
3. Test tube list. Displays the sample tubes used in the experiment. Right-click a tube in the list to perform additional
operations.
The test tube management module cannot add new sample tubes.
C44966AB 2-9
New Experiment Test Tube Management Module for DxFLEX [Without Autoloader] or DxFLEX [With
Autoloader - Single Tube Mode]
New Experiment Test Tube Management Module for DxFLEX [With Autoloader - Carousel Mode]
New Experiment Test Tube Management Module for DxFLEX [With Autoloader - Plate Mode]
Refer to Test Tubes in APPENDIX C, DxFLEX [With Autoloader -Plate Mode].
New Panel Experiment Test Tube Management Module for DxFLEX [Without Autoloader] or DxFLEX
[With Autoloader - Single Tube Mode]
New Panel Experiment Test Tube Management Module for DxFLEX [With Autoloader - Carousel Mode]
New Panel Experiment Test Tube Management Module for DxFLEX [With Autoloader - Plate
Mode]
Refer to Test Tubes in APPENDIX C, DxFLEX [With Autoloader -Plate Mode].
2-10 C44966AB
Plot area
1. Plot controls. For creating single or multiple plots, such as scatter plots, histograms, density plots, pseudocolor
plots, and contour plots.
2. Statistics and hierarchy controls. For creating statistical and hierarchical charts.
3. Graphical gating controls. For graphical gating of plots that have already been drawn.
4. Zoom controls. For zooming in and out.
5. Axis display controls. For scaling axis ranges in the plots.
6. Gain adjustment control. For increasing and lowering gain adjustments on the plots.
NOTE The gain adjustment control only works when a sample is running.
7. Adjust compensation control. For adjusting compensation of either of the parameters on a 2-D histogram.
8. Threshold control. For setting the minimum particle size limit or fluorescence intensity that acquisition will allow.
9. Undo and redo controls. For undoing or redoing an action in the drawing area.
10. Display controls. For controlling how plots and tables are aligned and arranged.
NOTE The Rearrange ( ) and Printing Controls ( ) icons can be accessed by selecting the
drop-down arrow at the end of the toolbar.
• Rearrange. For restoring the plots to the default positions.
• Printing controls. For printing and previewing the plot area.
11. Plot area. For drawing plots and displaying statistics and hierarchy tables.
C44966AB 2-11
Status Bar
1. Communication connection status. Displays whether the Cytometer and the Workstation are
connected.
2. Instrument status information. Displays the status of the Cytometer.
3. Sampler status. Displays the sample injection mode state. There are two sample injection modes:
Semi-automatic sample injection mode and manual sample injection mode.
NOTE DxFLEX Cytometers equipped with an autoloader have three sample injection modes: Single
Tube Mode, Carousel Mode, and Plate Mode. The Plate Mode requires a plate adapter.
4. Fluid status information. Displays the liquid level of the Fluid Containers.
Analysis Screen
The Analysis screen is similar to the Acquisition screen, without the acquisition control modules.
Drawing controls (see Figure 2.1) include the multi-data histograms and graphical display data
controls.
Figure 2.1 Drawing Controls Toolbar (Top of Screen)
2-12 C44966AB
Plots Zoom Threshold
Statistics Scale Undo/Redo
Hierarchy Gain
Gates Compensation
Compensation Experiment Screen

The Compensation Experiment screen appears when you open or create a new compensation
experiment.
1. Test tube management. Displays sample tubes required for the compensation experiment.
2. Plot area. Displays compensation plots and gating.
The test tube management section of the screen can import saved data (.fcs) files for computational
purposes.
Compensation Controls
The control area includes the compensation controls, coordinate axis display controls, gain
adjustment controls, and the undo and redo controls. The compensation controls give you the
option of calculating the compensation value, displaying the compensation matrix, or changing the
compensation parameters.
C44966AB 2-13
QC Experiment Screen
The Quality Control (QC) Experiment screen appears when you access a QC experiment.
QC Report Screen
Before starting the QC routine, a Settings screen appears.
Figure 2.2 QC Report Screen
1. Menu. Allows you to configure settings related to QC experiments.

2. Acquisition control. Controls sample loading/unloading and data recording.
3. Lot selection. Allows you to select the correct lot number of the QC reagent.
4. QC results list area. Displays the time and results of completed QC runs.
5. QC reports area. Displays detailed reports for the selected QC experiment.
2-14 C44966AB
QC Experiment Screen
When acquiring QC samples, the software opens the QC screen.
1. QC experiment progress indicator. Displays the QC stage.

2. Plot area. Displays the QC plots.
QC Screen Navigation
The Analysis screens have two navigation icons, one for the QC screen and the other for the
Levey-Jennings (LJ) plots.
c
1. QC screen icon. Accesses the QC screen.
2. LJ screen icon. Accesses the Levey-Jennings (LJ) screen.
C44966AB 2-15
Software Menu
The CytExpert for DxFLEX software contains the following selectable menu items:
Figure 2.3 Software Menu Tree†
† The menu options for File, Cytometer, Settings, and QC/Standardization change when you select Start
QC/Standardization. Refer to Figure 2.4.
*Boost is only active in the Manual Sample Injection Mode in the DxFLEX [Without Autoloader].
**Only available in Panel Experiments.
Figure 2.4 QC Software Menu Tree
*Boost is only active in the Manual Sample Injection Mode in the DxFLEX [Without Autoloader].
**Only available in Panel Experiments.
Acquisition and Analysis Screen Menu
2-16 C44966AB
File Menu
For creating new experiments, opening old experiments, saving new experiments and data, and
importing/exporting FCS data files.
C44966AB 2-17
Cytometer Menu
For configuring Cytometer settings and controlling Cytometer functions. Depending on the
Cytometer state, certain functions may not be available.
Standby state Initialized state
Settings Menu
Used to select and/or change software options and settings.
2-18 C44966AB
QC Menu
Select Start QC/Standardization from the QC/Standardization menu to start the QC or
Standardization routine.
Advanced Menu
Advanced settings are for experienced users and includes laser time delay settings.
Account Menu
Used to for user account management settings.
Log Menu
Used to access the User Management Operation Log or Maintenance Log.
Backup/Restore Menu
Used to backup and restore user log data.
C44966AB 2-19
User Management
Help Menu
For displaying software version information and system Instructions for Use.
User Management
IMPORTANT Only an Administrator can manage users.

User Management is used to create and manage user accounts.
Select Account > User Manager. The User Manager window appears.
Figure 2.5 User Manager (Card View)
2-20 C44966AB
User Management 2
Figure 2.6 User Manager (Grid View)
1. Search text box: Filters users by user name and 6. Unlock: Used to unlock an existing account that
full name. has been locked.
2. View drop-down: Toggles between Card View
(see Figure 2.5) and Grid View (see Figure 2.6).
NOTE An account locks after 3 failed password
attempts. The number of attempts can be
3. New: Used to create a new user profile. changed by the administrator. Refer to
4. Modify: Used to modify an existing user profile. Account Policies.
5. Delete: Used to delete an existing user profile.
NOTE An account automatically unlocks after
30 minutes. The duration can be changed
by the administrator. Refer to Account
Policies.
7. Reset Password: Used to reset an existing user

password to the default password: password.
8. Close: Closes the User Manager window.
C44966AB 2-21
User Management
Creating, Deleting, and Modifying Users in User Manager

Creating a New User in User Manager
1 Select in the User Manager window. The New window appears.
2 Fill in the new user information.

a. Enter the Username.
NOTE Username has the following naming requirements:

• The maximum number of allowable characters is 36.
• Special symbols are allowed.
• Username cannot be left blank.
b. Enter the Full Name.

NOTE Full Name has the following naming requirements:
c. Select the user Role.

d. Select the Enabled checkbox to enable the user.
NOTE The Enabled checkbox can only be changed by an administrator.
3 Select . The new user displays in User Manager.
4 Select .
2-22 C44966AB
User Management 2
Deleting Users in User Manager
IMPORTANT If an account has been used and log information has been generated related to it, the account
cannot be deleted, but it can be disabled.
1 Select the user to be deleted in the User Manager window then select .
NOTE The user 'Admin' is a system default user and cannot be deleted.
2 The following message appears: Are you sure to you want to delete the selected user? Select Yes to
confirm.
3 Select .
Modifying Users in User Manager
IMPORTANT If an account has been used and log information has been generated related to it, the
username cannot be modified.
1 Select in the User Manager window. The Modify window appears.
NOTE The user 'Admin' is a system default user and cannot be modified.
2 Modify the user information as necessary.
NOTE Uncheck the enabled box to disable a user.
3 Select .
C44966AB 2-23
User Management
4 Select .
Unlocking a User Account

Select a Locked user in the User Manager window and select Unlock.
NOTE You cannot unlock an active user.
Resetting a User Passwords

Select a user in the User Manager window then select . The user password is
automatically reset as password.
Changing a User Password
1 Select Account > Change Password. The Change Password window appears.
2 Enter the current password, the new password, and confirm the new password.
NOTE The new password must contain ten digits and all four of the following character types:
• letter in upper case
• letter in lower case
• numbers
• special character.
3 Select .
2-24 C44966AB
Role Management 2
Role Management

Role Management is used to manage user account permissions.
NOTE Multiple users can be applied to the same role.
Select Account > Role Manager. The Role Manager window appears. Refer to Figure 2.7.
Figure 2.7 Role Manager
1. New: Used to create a new role profile.

2. Modify: Used to modify an existing role profile.
3. Delete: Used to delete an existing role profile.
4. Close: Closes the Role Manager window.
C44966AB 2-25
Role Management
Creating, Deleting, and Modifying User Roles in Role Manager

Creating New User Roles in Role Manager
1 Select . The New window appears.
2 Fill in the new role information.

a. Enter the role name.
NOTE Role Name has the following naming requirements:

b. Enter the role description.
NOTE Role Description has the following naming requirements:

c. Select the permissions applicable to the new role.
2-26 C44966AB
Role Management 2
3 Select . The new role displays in the role list.
4 Select .
Deleting User Roles in Role Manager
IMPORTANT If a role has already been assigned to a user, that role cannot be deleted.
IMPORTANT The Administrator and Operator Roles are system defaults and may not be deleted.
1 Select the Role to be deleted in Role Manager then select .
2 Select .
Modifying User Roles in the Role Window
IMPORTANT The Administrator and Operator Roles are system defaults and may not be modified.
1 Select . The Modify window appears.
C44966AB 2-27
Account Policies
2 Modify the role information as necessary.
3 Select .
4 Select .
Account Policies

Account policies are used to define the default properties for the password policy, account lockout
policy, and application inactivity policy.
Select Account > Account Policies. The Account Policies window appears.
2-28 C44966AB
Account Policies 2
Figure 2.8 Account Policies - Password Policy
NOTE The allowable range for each entry is as follows:

• Password Recorded: 12-24
• Minimum Password Length: 10-12 characters
• Minimum Password Age: 0-89 days
• Maximum Password Age: 1-90 days
• Reminder for Expiration: 1-90 days
NOTE Minimum password age refers to a frozen period within which a password cannot be changed.
Maximum password age refers to a date when a password expires.
C44966AB 2-29
Account Policies
Figure 2.9 Account Policies - Account Lockout Policy

• Invalid Login Attempts: 3-5 times
• Lockout Time: 30-1,440 minutes
Figure 2.10 Account Policies - Application Inactivity Policy

• Inactivity Duration: 1-15 minutes
2-30 C44966AB
User Management Operation Log 2
User Management Operation Log
Viewing and Exporting User Logs
1 Select Log > User Management Operation Log. The Logs window appears.
2 Enter the filter conditions: User and Time Range.
3 To export the log, select .
NOTE User logs are exported as a .pdf file.
Maintenance Log
Beckman Coulter recommends maintaining the instrument periodically. Use the Maintenance Log
feature to archive, view, export, or manage the maintenance records. These records might be
requested by a service engineer for troubleshooting.
C44966AB 2-31
Maintenance Log
Adding/Deleting a Maintenance Entry

The maintenance entry is the information about the maintenance date and operator of a
maintenance task.
The Maintenance Log function tracks the maintenance information automatically for the following
tasks:
• System Startup Program

• QC
• Daily Clean
• Deep Clean
• Refilling Deep Clean Solution Bottle
• Replacing Peristaltic Pump Tubing
Follow the instructions below to add or delete the maintenance entry manually for the following
tasks:
• Clean Sheath Container

• Clean Waste Container
• Inspect the Liquid Flow Path for leaks
• Other additional tasks
1 Select Log > Maintenance Log. The Maintenance Log window appears.
2-32 C44966AB
Maintenance Log 2
2 Select a desired task and select Add Entry on the Maintenance Log window.
NOTE Add Entry is only available when the current date has been selected.
The checkmark displays on the screen indicating the maintenance entry has been recorded.
C44966AB 2-33
Maintenance Log
3 Optional: Select Delete Entry to delete a maintenance entry.

a. Select an entry, and select Delete Entry. The confirmation window appears.
a. Select Yes to confirm the deletion. The selected maintenance entry is permanently deleted.
Adding/Deleting Maintenance Tasks

In addition to the routine maintenance tasks, the users can add or delete a maintenance task
depending on their laboratory needs. The customized maintenance tasks display below the
Additional tab.
Adding a Maintenance Task
2-34 C44966AB
Maintenance Log 2
2 Select Add Task. The following window appears.
3 Enter a task name.
4 Select OK. The new task is added into the Additional Maintenance Task list.
C44966AB 2-35
Maintenance Log
5 Optional: Select Add Entry to record the maintenance information.
NOTE Add Entry is only available when the current day has been selected.
The maintenance information displays on the right.
6 Select Close.
2-36 C44966AB
Maintenance Log 2
IMPORTANT The routine maintenance tasks are set by default and cannot be changed or deleted from the
Maintenance Task list.
Deleting a Maintenance Task
IMPORTANT Take care not to delete a task that has maintenance entry records. Otherwise all the entries
of the current month will be permanently deleted.
2 Select a task to be deleted, and select Delete Task.
C44966AB 2-37
Maintenance Log
The confirm window appears.
3 Select Yes. The task is deleted from the Additional Maintenance Task list.
4 Select Close.
2-38 C44966AB
Maintenance Log 2
Viewing and Exporting Maintenance Logs

IMPORTANT The system allows you to share a workstation with different Cytometers. Make sure to select
the correct Cytometer Serial Number prior to viewing/exporting the maintenance log.
2 Optional: Verify that the Cytometer serial number displayed on the Maintenance Log window
matches with your Cytometer.
3 Select . The Export window appears.
4 Select the time range and select OK.
C44966AB 2-39
Maintenance Log
5 Select Save to export the file to a desired file path.
The maintenance log is exported.
2-40 C44966AB
Graphic and Gating Styles 2
6 Select Close to exit the Maintenance Log window. The maintenance log is exported.
Graphic and Gating Styles
Plots
The CytExpert for DxFLEX software offers a variety of plot formats including:
• Single-parameter plots and histogram overlays

• Dual-parameter plots: dot plots, density plots, pseudocolor plots, contour plots, and dot plot
overlays
NOTE Histogram Overlays and Dot Plot Overlays can only be created from multiple samples in the
Analysis or Report screen.
Histogram Multi-sample histogram overlays
Dot plot Density plot
C44966AB 2-41
Graphic and Gating Styles
Pseudocolor plot Contour plot Dot plot overlay
Gates
Various gating choices are available.
The software includes the following gate types:

• For dual-parameter plots: lasso, polygon, rectangle, four-quadrant, and hinged gates
• For single-parameter plots: line-segment and vertical gates
Lasso gate Polygon gatea Rectangle gate
Four-Quadrant gate Hinged gate Line Segment gatea
2-42 C44966AB
Software Settings 2
Vertical gate
a. This gate can be created using the autogate functionality. Refer to Creating and Adjusting Auto Gates in CHAPTER 6, Data
Acquisition and Sample Analysis
Software Settings
Select Options in the Settings menu to configure the software settings.
In the experiment settings, you can set the experiment’s default save path.
C44966AB 2-43
Software Settings
In the tube settings, you can select the columns that display in the test tubes section of the screen.
[Without Autoloader]
[With Autoloader]
2-44 C44966AB
Software Settings 2
In the plot settings, you can define the background of the graphics display area, configure the
histograms, and set the default signal parameters to either the channel’s area or the channel’s
height. The default is area. You can also set the default axis display range.
Opacity: 0-100
Coordinate axis default range:

Minimum: -16777215 to 16777215, Maximum: -16777215 to16777215 and greater than or equal to the
minimum value.
C44966AB 2-45
Software Settings
In the Gate settings, you can choose to display population percentage on all plots except overlay.
2-46 C44966AB
Software Settings 2
In the Page Setup settings, you can change the page size, orientation, margin size, and display
options.
Explanation of restrictions: Margin distance:

Minimum: 5.08
Maximum: May not cover current paper.
In the Carrier settings, you can set the following carousel settings:
C44966AB 2-47
Software Settings
Carousel Settings
• Check tube barcode
This function is to verify whether the inputted Sample ID information on the software matches
with the barcode label on the tube. Refer to Barcode Labels in CHAPTER 1, System Overview.
NOTE Refer to Software Settings in APPENDIX C, DxFLEX [With Autoloader -Plate Mode] for the plate
carrier settings.
NOTE When a mismatch is identified during an acquisition, the system stops the acquisition
immediately with a prompt message.
NOTE The Check Tube Barcode is active by default to prevent incorrect sample identification. When
the Check Tube Barcode is deselected, the Tube Barcode warning icon appears in the status bar.
• Mix and backflush settings
NOTE If the mix intensity is set to high, ensure the sample volume does not exceed 100μL. Sample
volumes that exceed this limit when the mix intensity is set to high could result in splashing.
Plate Settings
• Refer to Software Settings in APPENDIX C, DxFLEX [With Autoloader -Plate Mode] for the plate
settings.
2-48 C44966AB
Software Settings 2
In the Auto Export settings, you can choose to auto export reports and select the preferred file path,
export type and report type to be exported automatically.
Language Settings
Select Settings > Language Settings to open the Language Settings window. In the Language
Settings window, you can select which language to use for the software menus and graphical
statistics. The two options currently offered are English and Simplified Chinese.
C44966AB 2-49
Software Settings
2-50 C44966AB
CHAPTER 3
Operation Principles
Overview
This chapter explains how the Cytometer measures scattered light and fluorescence as cells pass
through the laser beam.
The illustrations in this chapter are not exact representations of the inside of the Cytometer. They
are for explanatory purposes only.
• Sample Flow
• Laser Beam Shaping
• Cell Illumination
• Light Collection, Separation and Measurement
• Signal Processing
• Data Storage
• Automated Software Features
• Parameters
• Plot Display
• Statistics
Sample Flow
CAUTION
Possible flow cell damage. To avoid clogging the sample probe, sample tubing or
flow cell, ensure that 12 x 75 mm test tubes are free of debris before you use
them.
Sample Loading
The sample carousel has bar-code labels that identify the carousel and the tube position number.
Also, you can put bar-code labels on the sample tubes. See APPENDIX B, Barcode Specifications for
the DxFLEX [With Autoloader].
C44966AB 3-1
Sample Flow
The Autoloader has a bar-code reader that reads the carousel number, the sample tube position, and
the sample tube bar-code labels before the carousel rotates and it loads sample tube. The
Autoloader handles a sample tube as follows:
• It lifts the tube out of the carousel using a centering cup.

• It moves the bottom of the tube in a circular orbit to mix the sample.
• It lowers its sample probe into the tube and the sample peristaltic pump starts to aspirate the
sample. Sample flow begins.
The sample probe is cleaned automatically when sample flow ends.
Hydrodynamic Focusing
The instrument uses a process called hydrodynamic focusing to ensure that the cells move through
the laser beam one at a time, along the same path through the flow cell.
The flow cell (Figure 3.1) contains a rectangular channel. A pressurized stream of sheath fluid
enters the channel at the lower end and flows upward. The sensing area of the flow cell is at the
center of the channel.
While the sheath stream is flowing through the channel, a stream of sample is injected into the
middle of the sheath stream. As shown in Figure 3.1, the sheath stream surrounds, but does not mix
with, the sample stream. The pressure of the sheath stream focuses the sample stream so that the
cells flow through the laser beam single file. If the cells were to move through the laser beam in
different ways during sample flow, sample analysis could be distorted.
3-2 C44966AB
Laser Beam Shaping 3
Figure 3.1 Flow Cell
e
d
b f
b
b
c
1. Sheath stream
2. Sample stream
3. Laser beam
4. Waste out
5. Purge port (Waste)
Laser Beam Shaping
Before the laser beam reaches the sample stream, lenses focus the beam (see Figure 3.2). Focusing
keeps the beam perpendicular to the sample stream flow while making the beam small enough to
illuminate only one cell at a time.
C44966AB 3-3
Cell Illumination
Figure 3.2 Laser Beam Shaping
1. Violet laser beam 4. First stage shaping lens

2. Blue laser beam 5. Second stage shaping lens
3. Red laser beam 6. Flow cell
Cell Illumination
As cells in the sample stream go through the sensing area of the flow cell, the elliptical beam
illuminates them. The cells scatter the laser light and emit fluorescent light from autofluorescence
and the fluorescent dyes attached to them.
Forward Scatter
The amount of laser light scattered at narrow angles to the axis of the laser beam is called forward
scatter (FS). The amount of FS is proportional to the size of the cell that scattered the laser light.
Side Scatter and Fluorescent Light

The amount of laser light scattered at about a 90° angle to the axis of the laser beam is called side
scatter (SS). The amount of SS is proportional to the granularity of the cell that scattered the laser
light. For example, SS is used to differentiate between lymphocytes, monocytes, and granulocytes.
In addition to the SS, the cells emit fluorescent light (FL) at all angles to the axis of the laser beam.
The instrument measures the amount of FL emitted by cells depending on the reagents used. For
example, FL above the background FL is used to identify molecules, such as cell surface antigens.
Light Collection, Separation and Measurement
Forward Scatter Collection

The FALS (Forward Angle Light Scatter) detector collects scattered light from a particle that
intersects with a laser and delivers information roughly proportional to the size of the particle. The
3-4 C44966AB
Light Collection, Separation and Measurement 3
forward angle light is filtered with a 488 nm band pass before it reaches the FS sensor which
generates voltage pulse signals. These signals are proportional to the amount of light the sensor
receives.
Side Scatter and Fluorescent Light Collection

Both side scatter and fluorescence are measured 90 degrees from the laser axis.
Side Scatter
The wavelength of SS is 488 nm. It is much more intense than FL.
Side scatter light collected by the objective lens is delivered by fiber optics to a patent-pending
design with high performance, solid-state, high efficiency, and low-noise detector arrays.
Fluorescent Light
Fluorescence and scattered light are transmitted by optical fibers to the Wavelength division
multiplexer (WDM). Each WDM is a unique detector array that corresponds to a different laser.
Refer to Wavelength Division Multiplexer (WDM) in CHAPTER 1, System Overview. Each WDM
contains optical filters and detectors for detecting channel fluorescence or scatter from a particular
laser. It is necessary to ensure that the filter and software settings match for each channel.
Figure 3.3 Light Path through the WDM with a Single Port
1. Fiber array photo detectors (FAPD)

2. Filter
3. 45-degree reflector
4. Doublet lens
5. Light path
6. Mirror
C44966AB 3-5
Signal Processing
Signal Processing
The DxFLEX is a fully digital system with an active range of 7 logarithmic decades, signal collection
speed of 30,000 events/second (includes 15 parameters).
Data Storage
CAUTION
The instrument Workstation is vulnerable to malware, viruses, data corruption,
and unauthorized instrument setting changes, or privacy breaches if
unauthorized access is gained by malicious personnel. To reduce the risk of such
events, only allow authorized laboratory personnel access to the instrument
Workstation. Contact your institutional security department for assistance.
CAUTION
The instrument Workstation is vulnerable to malware and viruses from physical
media such as CDs, DVDs, or USB drives. To reduce the risk of data corruption or
unauthorized setting changes, only use physical media such as CDs, DVDs, or USB
drives that are known to be free from viruses or malware. Contact your IT
professional for assistance.
Sample results can be printed out, saved to removable media, saved to a local hard drive or saved to
a network drive. You can store sample results in Flow Cytometry Standard (FCS 3.0) files.
Automated Software Features
CytExpert for DxFLEX software contains the following automated software features.
• Auto Delay. The system reads and automatically adjusts laser delay during QC. You can also read
the laser delay value while acquiring samples.
• Auto Threshold. Easily find target populations. No need to worry about the threshold setting
while adjusting gains when auto threshold is enabled.
• Auto Gating. There are two types of autogates available in the CytExpert for DxFLEX software:
auto line segment and auto polygon.
3-6 C44966AB
Parameters 3
Parameters
TIME Parameter
The TIME parameter is the amount of time, in seconds, the instrument acquires data. It is displayed
on the plot. The axis labels vary, depending on plot resolution and stop time (duration) if Fit with
sample is selected.
Figure 3.4 Time vs Fluorescence plot
Plot Display
The results of sample analysis appear on the Workstation screen as graphs called plots. Refer to
Graphic and Gating Styles in CHAPTER 2, Using the CytExpert for DxFLEX Software.
C44966AB 3-7
Statistics
Statistics
The Statistics Setting window allows you to change the display of the header, statistical elements
and cell populations included.
3-8 C44966AB
CHAPTER 4
Daily Startup
Overview
IMPORTANT Verify that the correct USB configuration key is securely connected to a computer USB port. If
the USB configuration key is not connected, the following error message appears: CytExpert cannot find
the license. Please check whether the correct USB configuration key has been plugged in.
This chapter describes the instrument startup procedure.

Workflow:
Pre-startup
Ô Turn on power Ô Open software Ô Initialize instrument
inspection

• Pre-Startup Inspection
• Turning On the Instrument
• Opening the Software
• Initializing the Instrument
Pre-Startup Inspection
Before using the DxFLEX flow cytometer, perform the following system checks.
C44966AB 4-1
Daily Startup
Pre-Startup Inspection
Check Waste and Reagent Levels
CAUTION
Risk of instrument damage. Do not use a saline-based sheath fluid on the DxFLEX
instrument. Saline-based sheath fluid could damage instrument components.
Beckman Coulter recommends using DxFLEX Sheath Fluid or a similar nonionic
strength sheath fluid to ensure system performance.
1 Examine the sheath fluid and waste containers. Verify that there is sufficient sheath fluid in the
sheath fluid container and that the waste container is empty.
NOTE When the sheath fluid container is near empty or the waste container is near full, a warning
message is transmitted to the Workstation and audible signals sound as a warning.
Fluid Containers [Without Autoloader Shown]
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1. Sheath fluid container

2. Waste container
CAUTION
Risk of instrument damage. Remove the sheath fluid container from the Fluid
Container holder before filling the sheath fluid container to avoid damage to
instrument electronics.
2 If necessary, fill the sheath fluid container with DxFLEX Sheath Fluid or a similar nonionic
strength sheath fluid while not exceeding the maximum volume indicated (4 L).
4-2 C44966AB
Daily Startup
Pre-Startup Inspection 4
WARNING
Risk of chemical injury from bleach. To avoid contact with the bleach, use barrier
protection, including protective eyewear, gloves, and suitable laboratory attire.
Refer to the Safety Data Sheet for details about chemical exposure before using
the chemical.
3 If necessary, empty all waste liquid from the waste container. If biohazardous samples are used
for data collection, add 400 mL of 5% to 6% bleach to the waste container.
4 Verify that the Fluid Containers and the Cytometer are on the same level.
5 Verify that all flow tubes and sensor cables are properly connected, as shown in the figure:
b c d
e f g
1. Waste level sensor connector. Connects to the waste liquid sensor cable.
2. Flow cell waste out. Connects to the flow cell waste tubing.
3. Sheath fluid level sensor connector. Connects to the sheath fluid sensor cable.
4. Waste out. Connects to the waste liquid tubing.
5. Sheath return. Connects to the sheath fluid tubing.
6. Sheath fluid in. Connects to the sheath fluid tubing.
C44966AB 4-3
Daily Startup
Turning On the Instrument
Power Source Inspection

Check the power cable located below the power switch on the back of the Cytometer, and verify it
is securely connected to both the Cytometer and the power source.
Workstation Connections Inspection

Check that the monitor, mouse, keyboard, and the Cytometer are properly connected to the
computer. Refer to Figure 1.15.
Turning On the Instrument
CAUTION
1. If the Cytometer or Workstation fails to start properly, check first to see
whether the power cable and connection cables are properly connected.
2. Never shut off the power or disconnect a data cable while the Cytometer is
performing a task. Doing so can result in data loss or damage to the system.
1 Turn on the main power switch located on the back cover of the Cytometer.
2 Wait for the Cytometer to finish powering on, then turn on the Workstation.
Opening the Software
The system performs an automatic system check when the software is opened.
1 Log in to the Windows operating system and select the CytExpert for DxFLEX desktop icon
to open the software.
4-4 C44966AB
Daily Startup
Opening the Software 4
The login window appears.
NOTE The default software shortcut appears on the desktop. If you do not see the icon, the default
installation path is under C:/Program Files/CytExpert for DxFLEX. Select CytExpert.exe to run the
software.
2 Enter your username and password.
3 Select .
NOTE The name of the user that is currently logged in displays in the top, right corner of the software
screen.
C44966AB 4-5
Daily Startup
4 Confirm that the software and the Cytometer are properly connected.
a. Open the software. The Startup screen appears.
b. Verify that the connection indicator light in the lower left corner of the software screen is
green, and Connected is displayed. The left side shows the connection status, the middle
shows the instrument status, and the right side shows the status details.
4-6 C44966AB
Daily Startup
c. Verify that the Sheath and Waste flow indicators in the lower right corner of the software
screen are green indicating that the fluidics system is normal.
NOTE
• If the connection indicator light is red, it means that there is a faulty connection. Ensure that
the instrument is properly turned on and connected. If necessary, restart both the
Cytometer and the Workstation.
• After the instrument initializes, a warning beep sounds if there is a problem with the fluidics
system. If a flow indicator is red and blinking, it means that the fluidics system requires
attention.
• When the waste liquid sensor is disconnected it shows that the waste container is full or
nearly full.
• Select the status information in the lower left to open the system log. Send a copy of the
system log to your Beckman Coulter Representative for support if a service call is
requested.
C44966AB 4-7
Daily Startup
Logging Out of the Software

Select the username displayed in the top-right corner of the software screen and select
.
Or select Lock to lock the software. The software locks automatically if it remains inactive for a
specified duration.
Selecting the Proper Sample Injection Mode [Without Autoloader]
Select Sample injection Mode in the Cytometer Menu to change between the Semi-Automatic
Injection mode and the Manual Injection mode. The Semi-Automatic Injection mode is
recommended under most circumstances. The Manual Injection mode can be used for two
4-8 C44966AB
Daily Startup
purposes: running 1.5-mL and 2-mL microcentrifuge sample tubes and a backup mode that allows
you to continue to collect data if the Semi-Automatic Injection mode is not working correctly.
Using Semi-Automatic Injection Mode
1 Select Sample Injection Mode > Semi-Automatic in the Cytometer menu to change the Sample
Injection mode selection. The sampler status icon located in the bottom right side of the screen
changes to display Semi-automatic Sampler.
2 Select Initialize. The sample tube holder swings out from the standby position to the sample
loading position (see Figure 1.11) so that you can load the sample tube.
NOTE You can also swing out the sample tube holder manually, load the sample tube, then select
Initialize.
3 Select Run. The sample tube holder automatically swings back to the standby position and
raises the sample tube to the sample acquisition position (see Figure 1.11), where the
instrument mixes the sample and transfers the sample to the flow cell.
C44966AB 4-9
Daily Startup
At the flow cell, the sample runs at the designated flow rate and the Cytometer begins to
acquire data.
NOTE You can also push the load button on the front of the instrument to automatically start the run
and record the data.
4 When you are satisfied with the appearance of the data, select Record to record the data.
5 Wait for the data acquisition to finish or select Stop. The sample tube holder automatically
lowers the sample tube and moves it to the sample loading position (see Figure 1.11) and the
Cytometer back flushes the sample probe.
WARNING
Risk of biohazardous contamination. When using 1.5-mL and/or 2-mL sample
tubes, always cut the cap off and do not exceed 300-μL sample volume. Running
samples with a cap attached to the sample tube or with volumes exceeding 300-μL
can result in sample splashing.
Using the Manual Injection Mode
1 Select Sample Injection Mode > Manual in the Cytometer menu to change the Sample Injection
mode selection. The sampler status icon located in the bottom right side of the screen changes
to display Manual Sampler.
4-10 C44966AB
Daily Startup
2 Manually swing the sample tube holder out from the standby position to the sample loading
position.
3 Select Initialize.
C44966AB 4-11
Daily Startup
4 Load the sample tube.
NOTE The sample tube holder accommodates 1.5-mL, 2.0-mL, and 12x75 mm sample tubes.
5 Manually swing the sample tube holder gently back to the standby position.
4-12 C44966AB
Daily Startup
6 Manually raise the sample tube holder gently to the sample acquisition position and hold the
tube in that position.
7 Select Boost to transfer the sample to the flow cell.
8 Select Run.
The sample runs at the designated flow rate and the Cytometer begins to acquire data.
9 When you are satisfied with the appearance of the data, select Record to record the data.
C44966AB 4-13
Daily Startup
10 Wait for the data acquisition to finish or select Stop. Then, manually lower the sample tube
holder and move it to the sample loading position.
11 Select Backflush to clean the probe.
Selecting the Proper Sample Injection Mode [with Autoloader]

Select Sample injection Mode in the Cytometer Menu to change between the Single Tube Mode,
Carousel Mode, and Plate Mode.
The Single Tube Mode can be used for running a single tube in a carousel. The Carousel Mode can
be used for running 12x75 mm sample tubes in a 32-tube carousel. Refer to Selecting the Plate
Sample Injection Mode [With Autoloader - Plate Mode] in APPENDIX C, DxFLEX [With Autoloader
-Plate Mode] for instructions on using the DxFLEX [With Autoloader-Plate Mode].
4-14 C44966AB
Daily Startup
[with Autoloader]
Using Single Tube Mode or Carousel Mode
1 Select Sample Injection Mode > Single Tube Mode or Carousel Mode in the Cytometer menu to
change the Sample Injection mode selection.
The sampler status icon located in the bottom right side of the screen changes to display Single
or Carousel.
C44966AB 4-15
Daily Startup
4 Install the carousel. Refer to Installing a Carousel in CHAPTER 1, System Overview.
5 If necessary, calibrate the carrier. Refer to Calibrating the Carrier [DxFLEX With Autoloader] in
CHAPTER 12, Replacement/Adjustment Procedures.
4-16 C44966AB
Daily Startup
Running the System Startup Program [Without Autoloader]

IMPORTANT Instructions on the software window varies depending on whether you are in semi-automatic
injection mode or manual injection mode.
If the fluidic self-check is enabled, the system startup program takes approximately 10 minutes. If
the fluidic self-check is not enabled, the system startup program takes approximately 8 minutes.
NOTE The fluidic self-check is an optional feature only available when the sheath damper upgrade is
implemented.
C44966AB 4-17
Daily Startup
2 Select System Startup Program in the Cytometer menu.
3 The System Startup Program window appears. Select Initialize.
System Startup Program Window in Semi-Automatic Injection Mode
4-18 C44966AB
Daily Startup
System Startup Program Window in Manual Injection Mode
4 Wait for the system to initialize. Follow the on screen software prompts, then select Start.
The instrument begins to prime or run the fluidic self-check. This process takes about 1 minute
if the fluidic self-check is not enabled. This process takes about 4 minutes if the fluidic
self-check is enabled.
[Fluidic Self-Check Enabled Shown]
C44966AB 4-19
Daily Startup
After priming or running the fluidic self-check, the system initializes again. The sample is
loaded automatically. This process takes about 3 minutes.
The sample tube is unloaded after sample acquisition has finished. The system uses the
remaining time to warm up.
4-20 C44966AB
Daily Startup
5 When warm up is finished, select Close to quit the startup program. The system is now
initialized.
Running the System Startup Program [With Autoloader]

IMPORTANT Instructions on the software window varies depending on whether you are in Single Tube,
Carousel, or Plate Mode. These instructions also vary depending on whether you are in semi-automatic
injection mode or manual injection mode. For instructions on running System Startup Program in Plate
Mode, refer to Running the System Startup Program [in Plate Mode] in APPENDIX C, DxFLEX [With
Autoloader -Plate Mode]. The Plate Mode is not validated for IVD use with the DuraClone B27 Reagent.
The system startup program takes approximately 10 minutes.
NOTE The fluidic self-check feature is always enabled by default on all DxFLEX [With Autoloader]
instruments.
C44966AB 4-21
Daily Startup
The System Startup Program window appears.
System Startup Program Window in Single Tube or Carousel Mode
4-22 C44966AB
Daily Startup
The carousel sets the tube location in position 1 by default. Only one well location can be
selected.
NOTE To deselect the tube, select the desired tube and select Set As Empty.
NOTE To select new deionized water tube location, select the desired tube and select Deionized Water.
The instrument begins to run the fluidic self-check. This process takes about 4 minutes.
System Startup Program Step 2 in Single Tube or Carousel Mode
After running the fluidic self-check, the system initializes again. The sample is loaded
automatically. This process takes about 3 minutes.
C44966AB 4-23
Daily Startup
The sample is unloaded after sample acquisition has finished. The system uses the remaining
time to warm up.
4-24 C44966AB
Daily Startup
C44966AB 4-25
Daily Startup
initialized.
Selecting Experiments from the Start Page

Refer to Start Page in CHAPTER 2, Using the CytExpert for DxFLEX Software.
4-26 C44966AB
Daily Startup
Initializing the Instrument 4
Initializing the Instrument
1 Select Initialize in the Data Acquisition Control screen or select Initialize in the Cytometer Menu
to initialize the instrument.
[Without Autoloader Shown]
NOTE [Without Autoloader]: If the instrument is in Semi-Automatic Injection mode during the
initialization process, the sample tube holder automatically shifts into the sample loading position
(see Figure 1.11).
2 Wait for the beep indicating that the instrument properly initialized.
NOTE After the instrument initializes, the laser powers on and the fluidics system begins to function.
In the initialized state, the laser powers on to achieve operating status, and the sheath fluid flows.
• If you need to execute a task with the Fluid Containers, do so with the instrument in standby
state.
• If the instrument remains idle for 10 minutes, the Cytometer automatically enters the standby
state.
NOTE After approximately 30 seconds, there should be a continuous flow of waste liquid from the
Cytometer to the waste container.
3 Proceed to the subsequent operations or select Standby to put the instrument in standby state.
[Without Autoloader Shown]
C44966AB 4-27
Daily Startup
Initializing the Instrument
4-28 C44966AB
CHAPTER 5
Instrument Quality Control and
Standardization
Overview
This chapter provides information on performing daily quality control (QC) on the DxFLEX flow
cytometer and how to confirm that the instrument is working properly within the specified
parameters. Quality control allows you to determine whether your instrument can provide
adequate signal strength and precision.
This chapter also provides information on performing standardization. DxFLEX Daily QC

Fluorospheres or any other reference material that is relevant for your applications may be used as
the standardization samples.
Standardization can be used to setup the instrument for any specific assay using the application
defined target values or the Median Fluorescent Intensities (MFI). Standardization, however, does
NOT replace QC as the Cytometer's optical alignment (rCV statistical analysis), Laser Power and
Laser Delay outputs are not measured during the run.
The QC process verifies important system functions. The system:

1. Verifies that the unit hardware configuration matches the default configuration specified in
the software. Refer to Verifying, Selecting, Editing, and Creating Detector Configuration in
CHAPTER 6, Data Acquisition and Sample Analysis.
2. Measures the laser power of each individual laser and ensures that each laser meets the system
specifications.
3. Loads the sample and begins to acquire data.
4. Verifies that the actual laser delays match those set in the software and will adjust the delay
within certain parameters.
5. Checks the laser delay settings.
• If laser delay is 2 μs from the previous setting, it is acceptable and the software does not
notify you.
• If laser delay is >2 μs but 5 μs from the previous setting, the software notifies you and
automatically changes the laser delay setting.
• If laser delay is >5 μs from the previous setting. Manual laser delay adjustments are
required. Refer to Setting Laser Delay in CHAPTER 12, Replacement/Adjustment
Procedures.
C44966AB 5-1
Instrument Quality Control and Standardization
Overview
6. Verifies and calibrates the gain settings. If any of these parameters are outside of the operating
limits, the system automatically adjusts these parameters. If the system is unable to adjust
these parameters to fall within the operating limits, the system notifies you.
NOTE Beckman Coulter recommends performing QC on a daily basis.
NOTE QC can only be run using the standard detector sets including, laser and band-pass. Refer to Verifying,
Selecting, Editing, and Creating Detector Configuration in CHAPTER 6, Data Acquisition and Sample
Analysis to verify that the default factory detector configuration is selected before running QC.
NOTE CytExpert for DxFLEX QC includes auto daily QC routine with Levey-Jennings (LJ) charts tracking and
logging.
NOTE CytExpert for DxFLEX standardization allows for application-specific settings to be applied to future
experiments.
QC Workflow:
Prepare the QC Select target

Ô Ô Acquire data Ô Confirm results Ô Daily clean
sample values
Standardization Workflow:
Obtain Run Generate Apply

Perform
acquisition Ô standardization Ô target Ô Ô standardization
standardization
settings beads medians settings
• Preparing the QC Sample Tube

• Importing Lot-Specific Target Values
• Collecting QC Data
• Confirming Results
• Standardization
— Generating Target Median Values
— Managing Standardization Target Library
— Performing the Standardization
— Applying the Standardized Acquisition Settings
5-2 C44966AB
Preparing the QC Sample Tube 5
Preparing the QC Sample Tube
Required materials
The following materials are required to complete the QC process:
• DxFLEX Daily QC Fluorospheres

• Sample tubes (12 x 75 mm).
• Vortexer
Preparation process
1 Take one sample tube and label it as the QC sample tube.
2 Use the vortexer or shake vigorously to thoroughly mix the bottle of DxFLEX Daily QC
Fluorospheres.
3 Add 10-15 drops of DxFLEX Daily QC Fluorospheres to the sample tube.
4 Use the vortexer to mix the sample tube until a uniform mixture is achieved.
5 Place the sample tube in the sample station.
Preparing the QC Well Plate
Refer to Preparing the QC Well Plate in APPENDIX C, DxFLEX [With Autoloader -Plate Mode].
C44966AB 5-3
Importing Lot-Specific Target Values
Import lot-specific target values for each new lot of DxFLEX Daily QC Fluorospheres.
CAUTION
Risk of erroneous QC results. Different target value information corresponds to
different lot numbers. Selecting the wrong lot number will lead to erroneous QC
results.
IMPORTANT Target gain values must be established for each new lot number of DxFLEX QC Fluorospheres.
QC could fail up to 3 times upon running each new lot number for the first time until target gain values
are established.
1 Select Start QC/Standardization from the QC/Standardization menu.
2 Select Target Library from the Settings menu. The Target Library window appears.
5-4 C44966AB
Importing Lot-Specific Target Values 5
IMPORTANT The Beckman Coulter website may prompt you to select your Region and Country prior to the
Beckman Coulter Technical Documents and Software page.
3 Select Download Target File. The Beckman Coulter Technical Documents and Software
Downloads page appears.
NOTE If your DxFLEX Workstation does not have access to the internet, navigate to
https://www.beckmancoulter.com/wsrportal/page/softwareDownloadSearch using a computer with
access to the internet and save the file to a USB drive. If the website is not accessible, contact us.
4 If necessary, register and log in to the Beckman Coulter website.
5 In the Search By Product section of the screen, select the following:

a. Select Research & Discovery from the Market Segment drop-down menu.
b. Select Clinical Flow Cytometry from the Product Line drop-down menu.
c. Select Instruments from the Product Series drop-down menu.
d. Select DxFLEX from the Product drop-down menu.
e. Select DxFLEX Daily QC Fluorospheres Target from the Software Name drop-down menu.
f. Select All from the Lot Number drop-down menu.
C44966AB 5-5
g. Select English from the Language drop-down menu.
6 Select Search.
7 The search results appear below the Search By Lot Number tab.
5-6 C44966AB
Importing Lot-Specific Target Values 5
8 Select DxFLEX Daily QC Fluorospheres Target Values under the Software Name column. The
DxFLEX Daily QC Fluorospheres Target Values page appears.
9 Select Download under the correct lot number from the DxFLEX QC Fluorospheres Target
Values page. The File Download pop up window appears.
10 Select Save to save the file to the desired file path.
11 Select Import from the Target Library window in the CytExpert for DxFLEX software.
C44966AB 5-7
Collecting QC Data
12 Navigate to the file saved in step 10 and select Open.
13 Select Close to exit the Target Library window.
Collecting QC Data
Before collecting QC data, be aware that QC data and reports are saved by default. To view or modify
the settings and file path, select QC/Standardization Setting in the Settings menu.
1 Double-click to start the CytExpert for DxFLEX software.

a. Ensure that the Connected icon on the Status Bar near the bottom-left side of the display is
green.
b. If the icon is not green, ensure that the Cytometer USB is securely connected to the
Workstation and restart the Workstation.
2 Verify the detector configuration. Refer to Verifying, Selecting, Editing, and Creating Detector
Configuration in CHAPTER 6, Data Acquisition and Sample Analysis.
NOTE Ensure that the instrument configuration is properly configured for the QC experiment. The QC
experiment may not be completed or may end in erroneous results if incorrect settings are chosen.
Beckman Coulter recommends using the factory configuration and ensuring that the proper optical
filters are in place.
5-8 C44966AB
Collecting QC Data 5
3 Select Start QC/Standardization in the QC/Standardization menu to access the QC experiment.
Ensure that the QC bead lot number is selectable in the Lot No. drop-down menu. If the lot
number is not selectable, refer to Importing Lot-Specific Target Values, then select the proper
lot number.
5 [Without Autoloader]: Insert the prepared QC sample tube (see Preparation process) into the
tube holder. Proceed to Step 7.
Or
[With Autoloader - Carousel Mode or Single Tube Mode]: Insert the prepared QC sample tube (see
Preparation process) into the carousel. Proceed to Step 6.
Or
[With Autoloader - Plate Mode]: Insert the prepared QC well plate (see Preparation process) into
the plate holder.
C44966AB 5-9
Collecting QC Data
IMPORTANT For instructions on setting up the plate, refer to Setting up the Plate in APPENDIX C, DxFLEX
[With Autoloader -Plate Mode].
6 Set up the carousel.
a. Select . The Carrier Settings window appears.
IMPORTANT Ensure the tube position on the carousel matches the tube position selected in the
software.
b. Select the sample tube location.

c. Verify mix and backflush settings.
d. Proceed to Step 7.
7 Select the lot number from the Lot No. dropdown.
NOTE If the correct lot number is not available, import target values. Refer to Importing Lot-Specific
Target Values.
NOTE Target gain values must be established for each new lot number of DxFLEX QC Fluorospheres. QC
could fail up to 3 times upon running each new lot number for the first time until target gain values
are established.
5-10 C44966AB
Collecting QC Data 5
8 Verify that Default is selected for Event Rate Setting. Refer to Changing the Event Rate Setting
in CHAPTER 12, Replacement/Adjustment Procedures.
If the Event Rate Setting is set to High, the following message appears.
9 Select Start to load the sample and begin to run the QC procedure.
The status of QC process appears on the left. Plots appear on the right. The QC experiment
sequentially detects configuration, laser power, laser delay, signal strength, and coefficient of
variation.
During QC, the software automatically seeks the DxFLEX Daily QC Fluorospheres and computes
the results. The software returns to the QC screen after the QC run is complete.
10 If the sampling rate is too low, the Cytometer stops the QC run and displays a prompt that the
QC run fails to reach the required event flow rate. This is not considered a QC failure. If this
situation occurs, increase the sample concentration by adding one drop of DxFLEX Daily QC
Fluorospheres to the sample tube and then perform the experiment.
C44966AB 5-11
Confirming Results
11 If the lot number of DxFLEX QC Fluorospheres is new and QC fails, the following software
message appears. Select Yes.
NOTE Target gain values must be established for each new lot number of DxFLEX QC Fluorospheres. QC
could fail up to 3 times upon running each new lot number for the first time until target gain values
are established.
If the lot number of DxFLEX QC Fluorospheres is not new and QC fails, refer to Step 3 or
CHAPTER 10, Troubleshooting.
If QC passes, proceed to Step 12.
12 Run Daily Clean to remove any residual fluorosphere particles. Refer to Daily Clean in
CHAPTER 11, Cleaning Procedures.
Confirming Results
Select Start QC/Standardization in the QC/Standardization menu to return to the QC Setting screen
to review Daily QC results.
1 Select the desired default configuration and date range from the drop-down menus located on
the left side of the QC screen to sort by the configuration used during the specified date range.
NOTE At least one date range must be specified.
5-12 C44966AB
Confirming Results 5
2 Select a QC run from the QC Process list on the left and a QC report appears on the right.
NOTE The results column indicates a passing QC result with a and a failed QC result with .
QC results must meet the following criteria to pass:
• The gain differences must be 20% from the target gain.
• The median fluorescence intensity (MFI) differences must be 5% from the target MFI.
• The rCV must be 5%.
C44966AB 5-13
Confirming Results
The report area on the right displays detailed experiment results, including laser power, delay,
testing conditions, and signal results. The same and symbols are used to indicate each
result. For items that fail, values falling outside the prescribed range are displayed in red font.
In the Comment area, an explanation appears for each failed item.
5-14 C44966AB
3 If QC fails, follow the procedure below:
NOTE Select No if the Confirm window appears.
a. Verify whether the beads used were within their shelf life and stored in accordance with
the appropriate instruction manual.
b. Verify whether the allocated sample tube was prepared as required and correctly
positioned.
c. Run Priming the Flow Cell in CHAPTER 12, Replacement/Adjustment Procedures, and
retest.
d. Run Daily Clean in CHAPTER 11, Cleaning Procedures, and retest.
e. Run Deep Clean Procedure in CHAPTER 11, Cleaning Procedures, and retest.
f. Repeat Steps c-d.
NOTE If QC fails two times in a row on the same day after repeating Steps a-f, contact us.
4 If necessary, you can select (for CSV format) or (for PDF format) in the top left corner
of the report area to export the QC results.
5 Select Close QC/Standardization in the File menu to exit the QC screen.
Creating Levey-Jennings Charts
1 Select Start QC/Standardization in the QC/Standardization menu to open the QC screen.
2 Select LJ chart on the left side of the screen.
C44966AB 5-15
Confirming Results
IMPORTANT When there are multiple plots, select which lot to create the LJ charts from.
3 Select LJ Chart Settings on the top of the LJ Chart screen. The LJ Chart Settings screen
appears.
5-16 C44966AB
4 Select the Laser tab, and select the power and/or delay check boxes for each laser as needed.
5 Select the Channels tab, and select each channel check box as needed.
C44966AB 5-17
Confirming Results
6 Select the Alarm Boundary and Scale Range tab, and set the acceptance criteria and the Y-axis
scale as needed.
NOTE “+/-2S” means the acceptance range is -2~2 standard deviations.
7 Select Apply.
8 Select OK.
5-18 C44966AB
9 Select the Levey-Jennings plot and select the start and end date from the drop-down boxes at
the top of the LJ Chart screen to specify the desired date range.
NOTE Select the desired configuration and date range from the drop-down menus located at the top of
the LJ Chart screen to sort by the configuration used during the specified date range.
10 Select Close QC/Standardization in the File menu to exit the QC screen.
QC Result Manager
The QC Result Management window can be used to search, delete, print, and export QC results.
C44966AB 5-19
Standardization
To access the QC Result Manager, right click the desired QC result and select Result Manager in the
QC screen. The QC Result Manager window appears.
Standardization
Use Beckman Coulter DxFLEX Daily QC Fluorospheres or any other reference material that is
relevant for your experiment. Ensure that either the Beckman Coulter DxFLEX Daily QC
Fluorospheres or other reference material has appropriate target values for your experiment.
NOTE Validation is required when applying the standardization target values to your application.
5-20 C44966AB
Standardization 5
Generating Target Median Values

IMPORTANT It is recommended to use Beckman Coulter DxFLEX Daily QC Fluorospheres for instrument
standardization. If you use other reference materials, follow your laboratory procedure to optimize the
gain settings for your experiment.
Follow the instructions below to apply the optimized gain settings to generate the target median
values, which will be applied to the standardization for your experiment.
1 Double-click to start the CytExpert software.

a. Ensure that the Connected icon on the Status Bar near the bottom-left side of the display is
green.
b. If the icon is not green, ensure that the Cytometer USB is securely connected to the
Workstation and restart the Workstation.
2 Select New Experiment to create an experiment.
3 Add a tube and edit the tube name. Refer to Changing the Tube Name in CHAPTER 6, Data
Acquisition and Sample Analysis.
4 Load the standardization beads.
5 Create a FSC/SSC dot plot and gate on the singlet bead population. Refer to Creating Plots and
Gates in CHAPTER 6, Data Acquisition and Sample Analysis.
6 Create histograms for each channel and apply the gate.
C44966AB 5-21
Standardization
7 Select Acq.Setting to input the optimized Gain and Threshold in the relevant channels in the
acquisition settings window.
NOTE The optimized Gain and Threshold settings are obtained using your laboratory procedure. Do not
adjust the Gain.
NOTE Use the FSC channel as the trigger channel. The threshold may need to be adjusted to visualize
the QC beads populations. If so, record this value for future reference.
8 Select Run.
NOTE If necessary, move the gate in the FSC/SSC plot to enclose the singlet bead population.
9 Select Record to save the experiment.
5-22 C44966AB
Standardization 5
10 Add a Statistics table and right-click the table and select Statistics Settings. The Statistics
Setting window appears.
C44966AB 5-23
Standardization
11 Select the Population tab and select the relevant population for the tube.
5-24 C44966AB
Standardization 5
12 Select the Statistics tab then select the Median Fluorescence value for all parameters used and
select OK.
NOTE The median values are the target settings that will be used for standardization.
13 Right-click the statistics table and select Export tube to CSV file.
If Excel is not available, manually record all the median values or take a screen shot.
C44966AB 5-25
Standardization
14 Save the experiment.

NOTE Rerun the experiment if:
• Standardization fluorosphere used are changed.
• The Lot number of standardization beads changes.
Managing Standardization Target Library

Select Standardization Target Library... from the Settings menu. The Standardization Target Library
window appears.
NOTE The Item name displays in the Acquisition Setting Catalog window as the saved acquisition setting
name.
NOTE Select to create a duplicate copy of an existing standardization item.
5-26 C44966AB
Standardization 5
Adding a New Standardization Item
1 Select . The Add Standardization Target Value window appears.
NOTE Unchecking the Use the same threshold setting for acquisition setting checkbox allows you to
specify custom threshold settings and when to save the test item into the Acquisition Setting
Catalog.
2 Enter the Item, Lot No., and Expire date from the drop-downs located at the top of the Add
Standardization Target Value window.
NOTE A single Lot No. can include several Items, but you cannot add duplicate Items under the same
Lot No.
NOTE If the Lot No. selected already exists, the Expire date cannot be edited.
3 Choose either Manual or Automatic threshold from the Standardization Test Setting or
Acquisition Setting section of the screen.
NOTE If you select Manual threshold, enter a value greater than 0, but less than 8,388,600.
C44966AB 5-27
Standardization
4 Set the channels, median, and median tolerance values.
NOTE The contents of the channel, laser and filter column come from the current detector configuration
setting.
NOTE Do not set the median tolerance range any lower than 5%.
NOTE FSC is a required channel.
5 Select OK to save the target value.

The saved results display in the Standardization Target Library window.
6 Select to exit the Standardization Target Library window.
Editing Standardization Item Parameters
1 Select an item from the Item column on the Standardization Target Library window and select
.
2 Edit the parameters for that item and select OK.
NOTE The Task Item, Lot No., and Expire date cannot be edited.
3 Ensure the item parameters are correct then select and save the file.
5-28 C44966AB
Standardization 5
Importing a Standardization Item
1 Select on the Standardization Target Library window.
2 Browse for the Item to import and select .

The imported item displays at the top of the list in the Standardization Target Library window.
Deleting Standardization Items

Select an item from the Item column on the Standardization Target Library window and select
. Select to exit the Standardization Target Library window.
Performing the Standardization
1 Open the CytExpert for DxFLEX QC screen.
2 Select the Standardization radio button.
[With Autoloader Shown]
C44966AB 5-29
Standardization
3 Select the Lot No. and the Items to be applied.
5-30 C44966AB
Standardization 5
4 Select .
The Process section of the screen displays the process details.
Once the process is complete, the Custom Setting Calibration Report displays.
C44966AB 5-31
Standardization
5 Verify the gain settings.

a. Select Acq. Setting Catalog from the Cytometer menu. The Acq. Setting Catalog window
appears.
NOTE designates test items from Standardization.
b. Select the desired Test Item, from the list in the Gain, Threshold, and Width tabs.
Applying the Standardized Acquisition Settings
1 Open an experiment.
NOTE The corresponding compensation matrix should have been determined as the proper settings.
Refer to CHAPTER 7, Compensation for detailed instructions on setting compensation.
5-32 C44966AB
Standardization 5
2 Select Acq.Setting from the Cytometer menu. The Acq. Setting window appears.
C44966AB 5-33
Standardization
3 Select Import from Catalog. The Acq. Setting Catalog window appears.
5-34 C44966AB
Standardization 5
4 Browse for the item to import and select Import.

The standardized settings are applied to the sample tube.
The Information window appears to notify of the corresponding channels with the changed
gain as a result of the Standardization.
C44966AB 5-35
Standardization
5 Select OK.
5-36 C44966AB
CHAPTER 6
Data Acquisition and Sample Analysis
Overview
This chapter contains information on how to use your DxFLEX flow cytometer, including data
acquisition, analyzing and exporting results, and compensation procedures that will be executed
manually during the process.
Workflow:
Adjust instrument
Create Load Record Analyze and Save
Ô Ô settings and Ô Ô Ô
experiment sample Data export data experiment
gates
• Creating Experiments
• Managing the Panel Library
• Load Sample and Record Data
• Configuring Acquisition Settings
• Analyzing and Exporting Data
• Saving the Experiment
Creating Experiments
There are two types of experiments available for use:
• Panel experiment: For creating a new experiment for a panel of tubes (multiple tubes grouped
together). The process creates a file with the .xitp extension and a folder with the same file
name where the raw data (.fcs files) are kept.
The panel experiment supports two worksheet modes: Independent worksheet mode and
Shared worksheet mode. Refer to Worksheet Modes.
• Experiment: For creating a new experiment for single tubes. The process creates a file with the
.xit extension and a folder with the same file name where the raw data (.fcs files) are kept.
The experiment only supports Independent worksheet mode. Refer to Worksheet Modes.
NOTE Experiments are also referred to as standard experiments in this section.
C44966AB 6-1
Creating an Experiment
1 Open the CytExpert for DxFLEX software and confirm that the instrument is connected. Refer
to Opening the Software in CHAPTER 4, Daily Startup.
Configuration.
3 Create or open an experiment using one of the following methods:

• Create a new experiment:
— Select New Experiment on the Start page, specify the file path, and save the experiment.
Or
— Select New Experiment in the File menu, specify the file path, and save the experiment.
• Create a new experiment from a template:

— Select New Experiment from Template on the Start page. Select Browse next to New
Experiment and specify the file path for the new experiment, then select Browse next
to Template and specify the file path to the existing template.
Or
— Select New Experiment from Template in the File menu, specify the file path and save
the experiment.
Or
— Select the Template tab on the Start page or select Recent Template in the File menu
and select the template from the list of recently used templates. Specify the file path
and save the experiment.
• Create a new panel experiment:

— Select New Panel Experiment on the Start page, specify the file path, and save the
experiment.
Or
— Select New Panel Experiment in the File menu, specify the file path, and save the
experiment.
• Open an existing experiment:

— Select Open Experiment on the Start page, specify the file path and save the
experiment.
Or
— Select Open Experiment in the File menu, specify the path and save the experiment.
Or
6-2 C44966AB
Creating Experiments 6
— Select the Experiment tab on the Start page or select Recent Experiment in the File
menu and select the experiment from the list of recently opened experiments. Specify
the file path and save the experiment.
NOTE Experiments are saved as a .xit file. Templates are saved as a .xitm file. Panels are saved as a
.xitp file. Panel experiments can also be saved as a template (.xitpm).
NOTE If you need to change the default save path, select Options in the Settings menu and modify the
Default Path displayed to the right of the Experiment tab. Then select OK.
Refer to Setting Tube Property [With Autoloader-Plate Mode] in APPENDIX C, DxFLEX [With
Autoloader -Plate Mode] for instructions on setting tube property using the Plate Mode.
Setting Tube Property [Single Tube or Carousel Mode]
1 [Standard Experiment]: Select to add tubes to the experiment. Skip to Step 3.
NOTE Use the dropdown to create multiple tubes.
Standard Experiment - Carousel Mode Shown
C44966AB 6-3
Or
[Panel Experiment]: Select to add samples to the experiment. Proceed to Step 2.
NOTE Use the dropdown to create multiple tubes or to duplicate existing tubes without data.
Panel Experiment - Carousel Mode Shown
2 [Panel Experiment]: Right-click each sample and select New Tube to add tubes to each sample.
NOTE To collapse all tubes and only view the samples, right-click a tube or sample and select Collapse
All. To expand all tubes, right-click a sample and select Expand All.
6-4 C44966AB
Samples Collapsed
Samples Expanded
3 Right-click each sample and select Edit Sample ID or double-click Sample ID box to input the
Sample ID information.
C44966AB 6-5
[Standard Experiment mode]:
[Standard Experiment]: Skip to Step 6.
[Panel Experiment mode]:
NOTE Ensure the Sample ID information matches with the barcode label if the tube has a barcode label.
Barcode labels are not required on sample tube. Refer to Barcode Labels in CHAPTER 1, System
Overview.
6-6 C44966AB
4 [Panel Experiment]: Right-click a sample then select Property. The Sample Property window
appears.
C44966AB 6-7
5 [Panel Experiment]: Set the sample properties for each sample.

a. Enter the sample type, collection time, submission time, submitted by, and test information
in the Sample Information tab of the Sample Property window.
b. Select the Patient Information tab.
6-8 C44966AB
c. Enter the patient name, patient type, patient ID, gender, D.O.B, age, charge type,
department, ward, bed number, clinical diagnosis, and attending doctor in the Patient
Information tab of the Sample Property window.
d. If desired, select File > Hide Personal and Sensitive Information to encrypt the submitted by,
reviewed by, test information, the patient name, patient type, patient ID, D.O.B, charge
type, ward, bed number, clinical diagnosis, and attending doctor.
C44966AB 6-9
e. The following message appears: The personal and sensitive information in the current
experiment will be replaced by “***”, are you sure you want to continue? Select Yes.
NOTE This encryption can not be retracted. Select No to cancel the encryption.
If you want to encrypt the personal and sensitive information only in the exported report, select
Hide Personal and Sensitive Information from the Batch Export Report window. Refer to
Batch Exporting Reports.
6-10 C44966AB
6 Right-click on a sample tube then select Property. The Tube Property window appears.
C44966AB 6-11
7 Set the tube properties for each tube. Enter the name, sample ID, and any desired remarks in
the Basic Information tab of the Tube Property window.
[Standard Experiment mode]
NOTE Sample ID is inactive in the Tube Property window if you are creating a new panel experiment. In
panel experiments, Sample ID is set in the Sample Property window. Refer to Steps 3.
8 Select . The Carrier window appears.
6-12 C44966AB
9 Select . The Add Carousel window appears.
10 Enter the Carousel number located on the Carousel barcode then select OK. The carousel image
appears on the Carrier window.
C44966AB 6-13
1. : Used to add a carousel.
2. : Used to delete the current carousel.

3. Carousel number drop-down menu: Used to toggle between carousels.
4. : Used to set tube locations in the carousel.
5. : Used to edit the backflush setting.
NOTE The default backflush setting is 3 seconds. You can choose to set different backflush
settings for each tube or press Control then select multiple tubes and set backflush for all
selected tubes.
6. : Used to edit the mix setting. Mix settings can be changed to an interval of X minutes, or
an interval of X wells. Mix duration and intensity can also be edited.
NOTE Mix settings vary depending on the sample injection mode selected. Single Tube Mode
does not have mix settings available. Carousel Mode only has mix duration available. Plate
Mode provides mix settings for mix duration, intensity, interval wells, and interval time.
NOTE The default mix setting is 3 seconds with an intensity of medium-high. You can choose
to set different mix time for each tube or press Control then select multiple tubes and set
mix time for all selected tubes.
7. : Used to clear the tube location.
8. : Used to set auto acquisition.
NOTE A number displays in the bottom right corner of each tube set for auto acquisition. For
example: .
9. : Used to cancel auto acquisition.
NOTE If a well is no longer set for auto acquisition, there is no number in the tube location. For
example: .
10. : Used to set the tube location for the cleaning agent.
11. : Used to set the tube location for the deionized water.
12. : Used to set the cleaning agent and deionized water acquisition time.
IMPORTANT When using Single Tube Mode, the tube location is automatically set to position 1 by default.
Skip to Step 15.
11 [Carousel Mode Only]: Select . The Set Location window appears.
6-14 C44966AB
Carousel Mode
12 Select the samples that should be added to the carousel.

NOTE can be used to select all samples. can be used to deselect all samples.
13 Select the tube location in the carousel or plate picture.

NOTE [Carousel Mode]: If more than one tube is selected, the samples will automatically group
together. To set individual well locations in separate locations (not grouped), select one tube at a
time.
14 Select OK.
NOTE To clear a tube or well location, select the checkbox next to the tube to be cleared then select
Clear Location.
15 Change the backflush and mix setting for each well or tube as needed.
NOTE Cleaning agent and deionized water tubes can only be added when using a carousel.
Worksheet Modes
There are two types of worksheet modes:
• Independent worksheet mode: One sample has multiple worksheets. Each tube in the sample has
its own worksheet. Independent worksheet mode can only be used with a panel experiment.
Each tube worksheet displays in the bottom portion of the screen. Refer to Figure 6.1.
C44966AB 6-15
Figure 6.1 Independent Worksheets
Refer to Switching to Shared Worksheets for instructions on how to switch from independent
worksheet mode to shared worksheet mode.
• Shared worksheet mode: One sample has one worksheet. Every tube in the sample shares the
worksheet. All items within the worksheet are shared with all tubes except the shape and
position of the gates and the statistics item within the statistics view. Shared worksheet mode
can be used with a standard experiment or a panel experiment. The shared worksheet displays
in the bottom portion of the screen. Refer to Figure 6.2.
Figure 6.2 Shared Worksheet
Refer to Switch to Independent Worksheets for instructions on how to switch from shared
worksheet mode to independent worksheet mode.
NOTE The worksheet display scale is fixed.
Switching to Shared Worksheets
1 Select in the bottom right corner of the screen. The Merge Worksheet window appears.
2 Select the reserve worksheet that all of the worksheets will be merged into.
3 Select OK.
Switch to Independent Worksheets
Select in the bottom right corner of the screen.
6-16 C44966AB
Changing the Tube Name

To change the name of a new sample tube or the sample ID, right-click the tube name or the sample
ID name in the Tube section of the screen and select Edit Name or simply double-click the sample
tube or sample ID name.
Setting the Channel and Label
1 Select Set Channel in the Settings menu. The Set Channel window appears.
C44966AB 6-17
2 In the Set Channel window, modify which channels are used and how they are displayed.
a. Select the channel signal check box, then you can add the reagent name in the Label
column. The information you add appears in the corresponding axis of the relevant plot in
the plot area. Unselected channel signals are not stored in the data file.
Panel Experiment Shown
b. Select Apply to. The Apply Channel Setting window appears.
c. Select the tubes to apply the channel settings to and select OK.
d. If you only need to modify the label name, select Set Label in the Settings menu to make the
required changes. The Set Label screen appears.
6-18 C44966AB
The Set Label screen does not allow you to select which channels to use, but it does allow
you to apply the modified label to all the sample tubes.
e. Select Apply to. The Apply Label Setting window appears.
f. Select the tubes to apply the label settings to and select OK.
C44966AB 6-19
Creating Plots and Gates

IMPORTANT The maximum number of elements allowed in a worksheet is 200. Elements include plots,
statistics tables, and gate hierarchy tables.
IMPORTANT The maximum number of gates allowed in a worksheet is 200.
1 Use the plotting controls (see Figure 2.1) in the plot area to create plots and gates and to
generate graphs as shown.
Use the icons to generate histograms, dot plots, density plots, pseudocolor plots, and
contour plot.
The experiment uses scatter plots, histograms, polygon gating, four-quadrant gating, and
line-segment gating.
a. After selecting a plot, click and drag the mouse to adjust the position and select and drag
the sizing handles at the edge of the graph to adjust the size of the graph.
6-20 C44966AB
b. Select an axis name to change which channel is displayed. An “A” after the channel name
indicates signal pulse area, while an “H” indicates height. The default setting is "A".
NOTE To modify the default settings, select Options in the Settings menu. The Options window
appears. Select Plot on the left side of the Options window. Under the Signal section of the
window, change the Main Channel default by selecting the Height or Area.
NOTE When using both Height and Area signals, ensure the gain setting is set to where the Height
signal does not reach its upper range.
C44966AB 6-21
c. Signal width can be used to differentiate somatic cell adhesion. If necessary, select
to open the Acq. Setting window.
6-22 C44966AB
d. Select the Width tab, and select a channel with the required signal width.
e. Plot properties can be configured to display axes in logarithmic, Log-Linear, or linear

format. To do so:
1) Right-click the plot and select Property from the drop-down menu. The Plot Property
screen appears.
C44966AB 6-23
Minimum: -16777215-16777215
Maximum: -16777215-16777215 and greater than or equal to the minimum value
Self-defining value: 1-16777215
2) Select whether to display the axes in logarithmic or linear format for both the X-axis
and Y-axis. Enter a value for log-linear coefficient if the log-linear view is desired.
3) Select Close.
Or
4) Select the logarithmic axis on the plot. The slider appears. Drag the slider along the
axis to change the log-linear coefficient and view events that are not shown, including
events with negative values.
NOTE The log-linear slider is also available during data acquisition.
NOTE To reset the axis back to logarithmic, right-click on the axis and select Property. Select
X axis Default or Y axis Default to reset the axis.
Histogram with logarithmic X-axis Histogram with log-linear X-axis
Dot plot with logarithmic X-axis Dot plot with log-linear X-axis
6-24 C44966AB
f. You can adjust axis ranges using the axis display controls located at the top of the screen.
• Select , to zoom-in and define which area of a plot to enlarge. The selected area
can be magnified to fill the entire graph. By selecting the zoom-out function, you can
click on the graph and restore the plot to its original appearance before magnification.
• Select to shift the axes. The mouse pointer appears as a hand. It allows you to
drag the graph to reveal the axis segment you need.
— Pan: Modifies the axis display range dimensions when panning both axes.
When the pan control is selected, you can right-click the graph and select which
axis you need to adjust when dragging. You can also pan directly to the default axis
range.
— Single side pan: Modifies the axis display range dimensions when panning one
axis.
NOTE Only the low end of the axis can be adjusted by the single side pan tool.
• Double-click the border area of the plot to open the Plot Property window, or
right-click the plot, then select Property to open the same Plot Property window.
• In the Plot Property window, manually enter the minimum and maximum display
values for the X- and Y-axes. You can also select Fit With Sample to let the software
automatically adjust the lower limit according to the signal and perform the
corresponding log-linear transformation. The X- and Y-axes Default settings are the
default parameters. The default parameters are 100-1,000,000.
NOTE Select Fit With Sample to identify the signal’s lower limit, adjusting automatically as
warranted. Selecting this item is recommended whenever the signal appears to be
relatively low.
NOTE Select Auto to automatically sets the upper and lower display limits of the axes based
on the data already collected.
NOTE Select Options in the Settings menu, then select Plot to modify the default setting of
the axis range under the Axis Default Setting section of the window.
C44966AB 6-25
2 To create gates, use the control buttons or right-click the

plot and select the gate type required. Gates can be set according to different requirements to
differentiate cell populations.
NOTE A newly created gate becomes a subset of the plot where it appears. The relationship between
parent and progeny/daughter gates can be changed when a displayed gate is subsequently
modified.
The position of the same gate in different sample tubes may vary. To change the position of a gate
and apply the change to all sample tubes accordingly, you can right-click the gate and select Apply
to All Tubes.
Use Link with to link multiple gates of a same type as a group. If you change the position or shape
of a gate on any plot, all the other gates will change at once accordingly.
6-26 C44966AB
a. Right-click the gate and select Link with to display the available gates.
b. Select the desired gate. The * displays on the gates as a link indication.
NOTE This Link feature is only available on the plots with same X and Y axis in a same worksheet in
acquisition mode.
C44966AB 6-27
3 Select the gates to display.

a. Select the heading area of the plot, select the parent population/gate to display in the plot
from the drop-down menu. The selected parent gate appears in the tab area of the plot.
NOTE The CytExpert for DxFLEX software will not list gates which would create circular gating logic.
Figure 6.3 shows all gates defined in the example experiment below. Note that the only gate
option in plot 1 of Figure 6.4 is P2 for the following reasons:
• Plot 1 cannot be gated on P1 because P1 is on that plot.
• Plot 1 cannot be gated on P2 because P2 is gated on P1.
• Plot 1 cannot be gated on the P2 OR P1 combo population because the gate logic contains
P1.
Figure 6.3 All Gates - Example Experiment
Figure 6.4 Circular Gating Logic - Example Experiment
6-28 C44966AB
b. If necessary, you can select the Combo Population option from the drop-down menu to
create a combination gate, using the Boolean relationships “and”, “or”, and “not” to
produce a new gate. You can also select the population color or change the gate name.
Name: The length of the string entered is less than or equal to 50.
• “And” indicates that all selections must be satisfied. For example, “P1 and P2” means
that the data for the newly added gate represent the intersection of P1 and P2.
• “Or” indicates that only one of the selections must be satisfied. For example, “P1 or P2”
means that the data for the newly added gate represent the union of P1 and P2.
• “Not” indicates exclusion from the selection. For example, “Not P1” means that the
data for the newly added gate represent the events that are not part of P1.
NOTE When creating a combo population gate, gates that are on the plot are not selectable.
C44966AB 6-29
4 Select to display the population hierarchy.

The Population Hierarchy function allows you to view how gates rank in relation to one
another. To change the display color, double-click the default color and select the desired color
from the drop-down color palette. To change the name of each gate, double-click the name of
the desired gate. By hovering your mouse pointer over a combo population whose display name
has just been changed, you can view its corresponding Boolean logical operation.
6-30 C44966AB
Creating and Adjusting Auto Gates

There are two types of auto gates available in the CytExpert for DxFLEX software: auto line segment
and auto polygon.
To create an auto line segment gate, select from the toolbar or right-click on the histogram
and select Auto Line Segment from the drop-down menu.
Select the population you want to gate in the histogram to automatically gate that population.
C44966AB 6-31
To create an auto polygon gate, select from the toolbar or right-click on the 2D plot and select
Auto Polygon from the drop-down menu.
Select the population you want to gate in the 2-D plot. The gate will automatically be drawn to fit
the population.
NOTE To add a vertex to an auto polygon gate:

1. Select the gate.
2. Hover your cursor over the perimeter of the gate until the cursor changes to the hand icon.
3. Select the desired location for the new gate vertex.
Turning Auto Recalculate On/Off

When auto recalculate is turned on, all autogates will recalculate when:
• The current tube is changed

• Compensation is changed
• Gating is changed
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• Collection stops
• An FCS file is imported to the tube or well
Auto recalculate turns off after a gate is moved or the size of a gate is altered. You must select Auto
Recalculate from the auto gate menu again to turn auto recalculate back on.
NOTE Auto recalculate turns on after adjusting movement or extent.
Right-click an autogate and select Auto Recalculate from the auto gate menu to toggle auto
recalculate on and off.
Adjusting Autogate Movement and Extent

Movement — The distance an autogate can move to find the target population.
To adjust movement, right-click an autogate and drag the Movement handle in the auto gate menu
left and right.
NOTE The default value setting for movement is 20 units. The minimum value setting for movement is 0
units and the maximum value setting for movement is 100 units.
If a target population is consistently in the same location, movement is not needed. However, if a
target population is periodically missing from some samples, or events are rare, movement can be
used to move the gate within a certain percentage of its axis to capture the correct population.
Refer to Figure 6.5 for an example of the default movement setting. Refer to Figure 6.6 for an
example of the maximum movement setting.
C44966AB 6-33
Figure 6.5 Movement - Default Setting
Figure 6.6 Movement - Max Setting
Extent — Shrinks or expands the gate around the population.

To adjust extent, right-click an autogate and drag the Extent handle in the auto gate menu left and
right.
NOTE The default value setting for extent is 20 units. The minimum value setting for extent is 0 units and
the maximum value setting for extent is 100 units.
6-34 C44966AB
Refer to Figure 6.7 for an example of the default extent setting. Refer to Figure 6.8 for an example
of the maximum extent setting.
Figure 6.7 Extent - Default Setting
Figure 6.8 Extent - Maximum Setting
Setting Customized Parameters

Set custom parameter to create fluorescence calculations.
1 Select Set Customized Parameter from the Settings menu. Or, right click a test tube from the test
tube menu and select Set Customized Parameter. The Set Customized Parameter window
appears.
Revised value: -999-9,999
C44966AB 6-35
2 Enter a name for the parameter in the Name section.
3 Select the parameters for calculation in the Parameter dropdowns.
4 Select the equation operations from the Oper dropdown menus.

The new parameter name displays in the list of parameters and statistic items.
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Setting Custom Statistics

Set custom statistics to create calculations based on populations of interest.
1 Right click the statistics table and select Statistics Setting. The Statistics Setting window
appears.
C44966AB 6-37
2 Select Expression.
3 Select Edit. The Expression window appears.
4 Enter the expression name in the Name section and enter the expression using the equation
buttons.
Name: String length may not exceed 30 characters.
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Managing the Panel Library 6
5 Select OK.
NOTE The equation populates in the Statistics Setting window under the Expression selection.
Managing the Panel Library
Select Settings > Panel Library to access the Panel Library window.
The Panel Library can be used to manage panels. Note that panels can be exported, imported, and
deleted from the Panel Library.
C44966AB 6-39
Figure 6.9 Panel Library Window
1. Panel list: Displays the list of panels available for use.

2. Import: Used to import panels.
3. Export: Used to export panels.
4. Delete: Used to delete panels.
5. Close: Used to close the Panel Library window.
Creating a New Panel Experiment

Refer to Creating Experiments for instructions on creating a new panel experiment.
Saving a Panel to the Panel Library
1 Right-click a sample in an existing panel experiment and select Export to Panel Library. The
Export to Panel Library window appears.
2 Enter the panel name and description.
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3 Select OK.
Importing a Panel
1 Select Settings > Panel Library. The Panel Library window appears.
2 Select the desired panel file.
3 Select Import.
4 Browse to the file location then select OK.
C44966AB 6-41
Exporting a Panel
2 Select the checkboxes next to the panels to be exported.
3 Select Export.
4 Browse to the file location then select OK.
6-42 C44966AB
Deleting a Panel
2 Select the panel or panels to delete.
3 Select Delete. The following system message appears:
4 Select Yes.
5 Select Close.
Using Heat Maps

Refer to Using Heat Maps in APPENDIX C, DxFLEX [With Autoloader -Plate Mode].
C44966AB 6-43
Load Sample and Record Data
Before Running Samples
CAUTION
Risk of erroneous results if the Cytometer has been idle for an extended period of
time. Perform a prime if the system has been idle for an extended period of time.
See Priming the Flow Cell in CHAPTER 12, Replacement/Adjustment Procedures.
1 Run the Daily Startup procedure.
2 Run the Instrument Quality Control and Standardization procedure.
3 Create an experiment. Refer to Creating Experiments.
4 Verify mixer settings. Refer to Changing Sample Mixing and Backflush Settings in CHAPTER 12,
Replacement/Adjustment Procedures.
5 Ensure that there is sufficient space on your hard drive for sample processing and data
acquisition.
Configuration.
7 Verify the laser settings. Refer to Laser Settings.
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Load Sample and Record Data 6
Verifying, Selecting, Editing, and Creating Detector Configuration
CAUTION
Risk of erroneous results. The system will read the selected Detector
Configuration even if the optical filters do not match the selected Detector
Configuration. You must verify the installed optical filters match the selected
Detector Configuration.
1 Select Detector Configuration in the Cytometer menu to verify the correct detector
configuration is selected. To change the configuration:
a. Select the desired configuration.
b. Select Set as Current.
A green checkmark appears in front of the selected configuration.
NOTE A configuration is locked when appears to the left of a configuration. A configuration locks for
two reasons:
• QC was run using the configuration.
• The compensation library contains data for the configuration.
Locked configurations can be deleted, but cannot be edited.
2 Select OK to close the Detector Configuration screen.
3 Proceed to Step 4 if you need to edit the Detector Configuration settings, or skip to Step 5 if you
need to create a new Detector Configuration, or skip to Step 12 if you need to delete a Detector
configuration.
C44966AB 6-45
4 If a saved configuration requires changes, edit that configuration.
NOTE The factory configuration is in bold and cannot be edited.
a. Select the configuration, then select Edit to access the Edit Detector Configuration screen.
b. Channels with a white background can be edited. Drag the names of the appropriate
fluorescence channels and optical filters on the left to the correct channels.
c. Continue to Step 6.
5 If an appropriate configuration is not saved, create a new configuration.

a. Select Detector Configuration... in the Cytometer menu.
b. Select New... and name your configuration.
You can also select a previously saved configuration and select Save As to create a copy.
c. Select OK.
6-46 C44966AB
d. Ensure the new configuration is highlighted, then select Edit. The Edit Detector
Configuration window appears.
e. Customize the new configuration. Channels with a white background can be edited. Drag
the names of the appropriate fluorescence channels and optical filters on the left to the
correct channels.
f. Continue to Step 6.
6 If a required channel name or filter is not listed on the left, select Fluorescence or Filter to add
or modify the channel name or the filter.
C44966AB 6-47
7 When finished, select OK.
8 Select the appropriate configuration.
9 Verify that the correct optical filters are installed in the Cytometer and match the newly
created configuration.
10 Select Set As Current.
11 Select OK.
12 To delete a configuration created in error, select Delete. The following confirmation message
appears. Select OK.
Sampling and Collecting Data
NOTE Settings can be imported from the Acquisition Settings Catalog. Refer to Importing and Exporting
Instrument Settings.
If compensation settings are desired, import the compensation from the Compensation Library or import
the compensation file. Refer to Importing and Exporting Compensation in CHAPTER 7, Compensation.
1 Select or from the Test Tube screen to create the new sample tube.
NOTE The first sample tube is already created by default in a standard experiment.
2 Change the tube name if necessary. Refer to Changing the Tube Name.
6-48 C44966AB
IMPORTANT Do not overfill the sample well.
3 Prepare and mix the sample intended for testing.
4 [DxFLEX Without Autoloader]: Ensure that the sample tube holder is in the sample loading
position. If the sample tube holder is not in the sample loading position, select Initialize.
C44966AB 6-49
WARNING
Risk of biohazardous contamination. When using 1.5 mL and/or 2 mL sample
tubes, always cut the cap off and do not exceed 300-μL sample volume. Running
samples with a cap attached to the sample tube or with volumes exceeding 300-μL
can result in sample splashing.
5 [DxFLEX Without Autoloader]: Place the sample tube in the sample tube holder.
IMPORTANT Ensure the Sample Injection Mode is set to the correct mode. Refer to Selecting the Proper
Sample Injection Mode [with Autoloader] in CHAPTER 4, Daily Startup.
[With Autoloader-Carousel Mode]: Place the sample tube in the carousel and then place the
carousel in the autoloader. Refer to Installing a Carousel in CHAPTER 1, System Overview.
[With Autoloader-Plate Mode]: Place the well plate in the plate adapter and then place plate
adapter in the autoloader. Refer to Installing the Plate Adapter in APPENDIX C, DxFLEX [With
Autoloader -Plate Mode].
6 Select the desired acquisition parameters (Events/Time to Record and Sample Flow Rate) on
the left side of the screen.
6-50 C44966AB
7 Select Run to load the sample.
NOTE When you select a tube that contains acquired data, as indicated by the blue tube in the test
tube section of the screen, the following message appears:
• Create new tube. Saves the current tube and creates an additional tube.
• Overwrite the data. Overwrites the current tube data with new data.
• Reacquire the data. Reacquires the data.
C44966AB 6-51
NOTE When you select a tube that contains recorded data, as indicated by the green tube in the
test tube section of the screen, the following message appears:
• Create new tube. Saves the current tube and creates an additional tube.
NOTE If you are running a DxFLEX [Without Autoloader] or a DxFLEX [With Autoloader - Single Tube
Mode], you can also push the load button on the front of the instrument to automatically start the
run and record the data.
8 View the plots and establish the gates. Refer to Creating Plots and Gates. Adjust the gate and
instrument settings as necessary. Refer to Configuring Acquisition Settings.
9 Adjust the gain settings. Refer to Adjusting the Gain.
10 Adjust the threshold settings. Refer to Adjusting the Threshold.
11 Adjust the Acquisition conditions. Refer to Setting Collection Conditions.
6-52 C44966AB
12 Select Record to save the data.

Wait for the saving process to finish. The sample tube holder returns to the sample loading
position (see Figure 1.11).
NOTE When you select a tube that contains recorded data, as indicated by the green tube in the
• Create new tube. Creates a new tube in the test tube section of the screen for the data.
• Append data to existing file. Adds new data to the existing data.
NOTE When you select a tube that only contains acquired data, as indicated by the blue tube in the
• Create new tube. Creates a new tube in the test tube section of the screen for the data.
• Overwrite the data. Overwrites the current tube data with new data.
• Append data to existing file. Adds new data to the existing data.
C44966AB 6-53
Configuring Acquisition Settings
Events to record: 1-100,000,000
Time to record: ≤1800 seconds
13 Repeat steps 1-12 until all sample tube data required for testing has been collected.
NOTE If the rate suddenly appears to drop, check to see if the sample has run dry or the sample probe
is clogged. Any time the sample probe becomes clogged, immediately select Stop to unload the
sample. Then select Backflush to clean the sample probe. Refer to Daily Clean in CHAPTER 11,
Cleaning Procedures, to flush out the sample probe. If you are still unable to clear the sample probe,
contact us.
Laser Settings
To access the Laser Setting window, select Advanced > Laser Setting. The Laser Setting window
appears. Refer to Figure 6.10.
NOTE The instrument must be in Standby mode to access the Laser Setting window.
Figure 6.10 Laser Setting Window
1. Enable/Disable: Enables or disables the laser.
IMPORTANT The Actual Power readings are +/- 1 mW.

2. Target Power: This function is not available on the DxFLEX.
3. Set: This function is not available on the DxFLEX.
4. : Reads the current laser power before the flow cell assembly and displays the current
laser power in the Actual Power (mW) column of the Laser Setting window.
6-54 C44966AB
Configuring Acquisition Settings 6
Select the Enable or Disable radio button next to each laser on the Laser Setting window to enable
or disable lasers. The laser status for each laser displays in the software status bar. Hover your
mouse over to display details for each laser.
NOTE Lasers can only be enabled and disabled when the system is in standby mode.
Adjusting the Gain

While the instrument is in use, the signal value can be increased or decreased by adjusting the
instrument's gain configuration.
1 Select on the left side of the screen. The Acq. Setting window appears.
2 Select the Gain tab in the Acq. Setting window.

Select or edit the instrument’s default gain settings using one of the following methods:
• Edit the gain settings and select Set as Default to create a new default setting.
• Select Default to return to your saved default settings.
NOTE In cases where you do not specify your own default parameters, the recommended settings and
default settings are identical.
C44966AB 6-55
3 Adjust the gain setting of each channel under the Gain tab in the Acq. Setting window. Raising
the gain increases the signal. Lowering the gain reduces the signal.
Gain range: 1-3,000
Another option is to use the Gain Control button on the toolbar in the graphic control area
to adjust the gain values for cell population data to their desired levels, directly on the plots
where the data appears during data collection.
NOTE Gain adjustments have a predefined range between 1 and 3,000. For fine adjustments, use the
text box under the Gain tab in the Acq.Setting window.
4 If necessary, change the coordinate display range and the plot type.
Adjusting the Threshold

By adjusting the threshold, the user can remove unnecessary signal noise to ensure that most of the
data collected consists of desired signal data. After the threshold settings have been configured for
a given channel, the acquisition of data from this channel will only be triggered by signals that
6-56 C44966AB
exceed the established threshold. Threshold settings have considerable bearing on whether the
appropriate events can be acquired.
1 Create a plot to view the channels where the threshold will occur. Generally, a bivariate plot
showing FSC and SSC is used.
NOTE Threshold can be defined for any of the fluorescence channels.
2 Select on the left side of the screen.
3 Select the Threshold tab in the Acq. Setting window.
Threshold: Manual: 1-8,388,600
NOTE It is recommended to select Height for Manual mode.
C44966AB 6-57
4 Set the desired threshold using one of the following methods:

• Choose the channel that is used for setting the threshold. Manually enter the threshold
value in the Threshold tab.
NOTE For dual-parameter plots, you can right-click the plot and select both parameters if desired.
Then, select the desired threshold boundary for the second parameter.
• Select Automatic in the Primary Threshold Trigger Level section of the Acq. Setting screen
to seek the target signal based on the background signal. It can quickly help find the target
population if the signal-to-noise ratio (SNR) of the channel is comparatively good. The
threshold can be set to either “H” (signal height) or “A” (signal area).
NOTE The automatic threshold value is based on the relative signal difference. When adjusting
gain, you do not need to update the threshold settings. For channels with a low SNR or an
excessively impure signal, manually setting the threshold parameters is recommended.
NOTE It is recommended to use Manual mode to set the threshold parameters to obtain accurate
experiment data.
Moreover, “and” as well as “or” can be applied to as many as two channels, so as to allow
these Boolean logical operators to be used in setting the threshold value.
— “and”: Data is displayed and collected only when two threshold conditions are met
simultaneously.
— “or”: Data is displayed and collected when at least one of two threshold conditions are
met.
• Select from the plot control area. Move your mouse pointer to the desired threshold
position in the desired plot and select once.
5 Select Close to start using the new threshold settings.
Setting Collection Conditions
1 Check mark the conditions required to set the necessary stop count events on the left side of
the Acquisition screen.
Two stop count collection conditions are available for sample recording:
• Events to Record. Used to set the number of events to record in the specified population.
• Time to Record. Used to set the collection time duration in seconds.
6-58 C44966AB
For example, if the event to record is set to record 1,000 P1 events, the software automatically
stops recording when P1 events reach 1,000 events. However, the software saves all data
acquired, including events outside of P1, when 1,000 P1 events is reached. You can also specify
the time to store if necessary. When multiple acquisition conditions are established, any one of
these conditions stops the collection process.
2 Select Record and wait for the software to complete collecting the data, at which time the
sample tube holder returns to the sample loading position (see Figure 1.11).
C44966AB 6-59
3 If you made changes to the data acquisition conditions and need to apply these changes to an
established sample tube, right-click the sample tube and select Apply Acq. Settings To, to apply
the conditions accordingly.
Setting Plot Display Conditions

Select Events Display Setting in the Settings menu. The Events Display Setting window appears.
6-60 C44966AB
Analyzing and Exporting Data 6
Three display options are available:
• Display all events. Used to view all events on the plot.

• Display first XXXX events. Used to set the set number of events to display.
NOTE The selected number of events displays in the bottom, left corner of the plot. For example, if you
choose to show 5000 events, the bottom, left corner of the plot displays 5000 Show.
• Display XX percent of events acquired. Used to set the percentage of events to display.
NOTE The selected percentage of events displays in the bottom, left corner of the plot. For example, if
you choose to show 20 percent of events acquired, the bottom, left corner of the plot displays 20%
Show.
Analyzing and Exporting Data
1 Select the sample tube to be analyzed.
C44966AB 6-61
2 Establish new gates or adjust the position of existing gates. Refer to Creating Plots and Gates.
NOTE Changing a gate’s position does not affect the positions of other gates already established on a
given sample tube. Each test tube individually records the positions of its associated gates. If you
need to make a change that concerns all the tubes, you must select the gate, then right-click the
correctly positioned gate and select Apply to All Tubes.
3 Select . The Gate Hierarchy screen appears.
4 Check the relationship between the parent and daughter gates in the Gate Hierarchy window.
NOTE Newly added gates become subsets of populations displayed in plots with existing gates. Name
and display color can be modified. Right-click directly on a gate plot to change the name and color.
NOTE Select No Color to leave the gated events uncolored but retain the color of their parent
populations. By default, the populations defined by vertical gate, hinged gate, or four-quadrant gate
are uncolored.
6-62 C44966AB
5 Right-click the plot and select Bring population to front to make the display color of the specified
gate appear in front of all other colors, or select Send population to back to hide the display
color of the specified gate behind all other colors.
6 Select in the plot area to generate a statistical table.
7 Right-click the table and select Statistics Setting to modify the settings of the statistics display
parameters. The Statistics Setting window appears.
C44966AB 6-63
The Statistics Setting window allows you to change the display of the header, statistical
elements and cell populations included.
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The final generated plots appear as below.
8 Right-click a plot and select Export to Graphic File or Export Tubes to Graphic File from the
drop-down menu to select an image to export.
• Export to Clipboard copies the plot to the clipboard, allowing you to paste it directly into
documents in common file formats.
NOTE Multiple plots can only be copied and pasted into Microsoft® Word. If a single plot is copied,
this can be pasted into both Microsoft® Word or Microsoft® PowerPoint.
• Export to File saves the plot as an image file.
NOTE Export to File can export plots in two selectable file formats. BMP bitmap format and EMF
vector format.
C44966AB 6-65
9 To export statistics, right-click a statistical table to select any one of the available export
options.
• Export to File exports individual tube statistics as a single CSV file.

• Export All Samples to File exports all tube statistics as a single CSV file.
• Export to Clipboard copies the statistics of the samples to the clipboard, allowing you to
paste them directly into a Microsoft® Excel file or other file formats.
• Export All Samples to Clipboard assembles the statistics for all the sample tubes of an
experiment and copies them together to the clipboard. From there they can be pasted as a
group into a Microsoft® Excel file or other file formats.
• Copy converts a statistical table into an image format that can be pasted into documents.
10 Export the FCS file if necessary. Refer to Exporting FCS Files.

NOTE Ensure that any storage devices used with the instrument are free from viruses. To guard against
data loss, Beckman Coulter recommends backing up data on a frequent and regular basis. Beckman
Coulter is not liable for any loss of data resulting from computer viruses or damage to hardware.
6-66 C44966AB
Exporting FCS Files

Exporting Single Tube Files
1 Right click the desired tube from the test tube section of the screen and select Export FCS File.
The Export FCS File window appears.
C44966AB 6-67
2 Select the population from the Population dropdown menu.
3 Select either Area or Height.
4 Select the FCS format next to File Version.
NOTE The default setting is FCS 3.0. If FCS 2.0 is selected, select the parameter type (linear or log) from
the parameter type section of the window.
NOTE If high auto-fluorescence vector values are present, the data is displayed differently in third party
software packages than in CytExpert for DxFLEX. Use the FCS 3.0 (High auto-fluorescence) option if
your file has high autofluorescence vector values. Auto-fluorescence values are added for the FCS
3.0 (High auto-fluorescence) option to accommodate the use of third party software. Since both FCS
3.0 options have the same .fcs file extension, ensure that you save the FCS 3.0 (High
auto-fluorescence) files to a different folder than the FCS 3.0 files.
5 Select the path to save the FCS file to from the Path section of the window.
6 Select Next to export the file.
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Exporting Multiple FCS Files
1 Select Export FCS File from the File menu. The Export FCS File window appears.
2 Select the tubes to export.
3 Repeat Steps 2-6 from Exporting Single Tube Files.
C44966AB 6-69
Exporting Plots or the Statistics Table of Multiple Tubes as Picture Files
1 Select File > Export FCS File. The Export Tubes to Files window appears.
2 Select the tubes to export and select Next.
3 Select the path.
4 Select OK.
NOTE The plots of the selected tubes save as .bmp file.
Batch Exporting Reports

Use the batch export report function to export the reports for selected sample tubes to a specified
directory. Reports can be exported as a .pdf, .csv, or image file.
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1 Select File > Batch Export Report. The Batch Export window appears.
2 Select the tubes to export from the Tube Name section of the screen.
3 Select the desired export options.
4 Select the desired export path.
C44966AB 6-71
5 Select Hide Personal and Sensitive Information to encrypt the personal and sensitive
information of a sample. The submitted by, reviewed by, and test information, patient name,
patient type, patient ID, D.O.B, charge type, ward, bed number, clinical diagnosis, and attending
doctor display as *** in the exported report.
6 Select OK.
Automatically Exporting Reports

NOTE This feature is only available when Auto Record is selected in Carousel mode.
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1 Select Settings > Options. The Options window appears.
2 Select the Auto Export tab.
C44966AB 6-73
3 Select Auto Export Report checkbox.
4 Select a default file path.
5 Select the export type.
6 Select Default Report from the Export Report section to export all default reports.
OR
Select All Report from the Export Report section to export all reports.
7 Select OK.
Importing and Exporting Instrument Settings

The CytExpert for DxFLEX software supports importing and exporting instrument settings to
facilitate the experiment process. Only instrument settings identical to the current configuration
can be imported with current detector settings.
Select to edit gain, threshold, and width. These can be imported from an experiment
file or from a catalog of instrument settings.
Importing Instrument Settings
1 Select the desired sample tube to import. Then select .
NOTE Instrument settings can only be imported into tubes where data has not yet been recorded.
6-74 C44966AB
2 Select Import From File, locate the file with the required instrument settings, or select Import
From Catalog to import the instrument settings.
3 Select Close.
Exporting Instrument Settings
1 Select the desired sample tube to export. Then select .
2 Select Export To File to export a current set of instrument settings, stored in a file ending in .acq.
Or
Select Export To Catalog, give a name to the settings to be exported, and export the file to the
software’s Acquisition Setting Catalog, then select OK.
3 Select Close.
Importing and Exporting Compensation Settings

The software supports unrestricted importing and exporting of compensation data, regardless of
whether the sample tube data has already been acquired. Imported compensation values only cover
channels identical with the current instrument configuration. The software automatically adjusts
C44966AB 6-75
compensation values according to differences in the gain level. Refer to Importing and Exporting
Compensation in CHAPTER 7, Compensation.
Printing Graphics
CytExpert for DxFLEX offers printing functionality for the plots and tables that appear in the plot
area. The software also lets you save these images by converting them into .jpg or .pdf files.
Select in the printer control area to print directly. Or, select the print drop-down arrow for
the following options:
• Print Preview. Used to access the Preview screen.
— Select to select the required format of the file to be exported and to save the file in
that format.
6-76 C44966AB
— Print preview also lets you choose between printing directly (1), modifying the printer
configuration (2), or adjusting the page settings (3).
b c d
• Page Setup. Used to adjust the page settings.
• Batch Print. Used to print data for multiple tubes.

1. Select Batch Print. The Batch Print window appears.
2. Select the tubes to print.

3. Select OK.
• Batch Export to PDF File. Used to print a PDF of the data for multiple tubes.
1. Select Batch Export to PDF File. The Batch Print to PDF File window appears.
C44966AB 6-77
2. Select the tube to print to PDF.

3. Select OK.
Printing Batch Reports

Use the batch print report function to print the reports for selected sample tubes.
1 Select File > Batch Print Report. The Batch Print window appears.
2 Select the tubes to export from the Tube Name section of the screen.
3 Select the desired export option.
4 Select OK.
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Saving the Experiment 6
Saving the Experiment
Selecting Save in the File menu allows you to save the experiment.
Selecting Save As and saving the experiment under a different name allows you to create a backup.
Selecting Save As Template in the File menu allows you to save the experiment as a template.
Concluding the Experiment

Conclude the experiment as follows:
• Select Standby to return the instrument to the standby state.

• Shut down the system. Refer to CHAPTER 9, Daily Shutdown.
C44966AB 6-79
Saving the Experiment
6-80 C44966AB
CHAPTER 7
Compensation
Overview
This chapter describes how to create a compensation experiment and automatically calculate
compensation values after acquiring the data. It also explains how to use these calculations for
other experiments.
Compensation involves correction for fluorescence spillover emitted by the primary fluorochrome
that is detected by the secondary fluorescent channels. For example, the excitation and the
resulting fluorescence emission for the PE fluorochrome leads to the spillover fluorescence
detected in the ECD, PC5.5, and PC7 channel. Compensation reduces the spillover fluorescence of
the PE-positive population to match the background of the PE-negative population in the secondary
channels. Compensation requires a single positive and a negative population for every single color
sample.
Properly configured compensation eliminates false data interpretation caused by spillover

fluorescence from another fluorochrome. Refer to Figure 7.1 and Figure 7.2 for an example of plots
C44966AB 7-1
Compensation
Overview
before and after compensation. Compensation adjustments can be completed during the data
acquisition process or after the data acquisition process is complete.
Figure 7.1 Before Compensation
Figure 7.2 After Compensation
NOTE CytExpert for DxFLEX compensation allows full matrix compensation, manual, and automatic.
CytExpert for DxFLEX compensation also includes a novel Compensation Library for storage of spillover
values of dyes to easily determine the correct compensation matrix with new gain settings.
NOTE Compensation is not validated for IVD use with the DuraClone B27 Reagent.
Workflow:
Apply
Prepare Create
Acquire and Calculate compensation
Compensation Ô compensation Ô Ô Ô
record data compensation to sample for
Samples experiment
data analysis
• Creating a Compensation Experiment

• Creating the Compensation Matrix from Previously Acquired Data
• Adjusting Compensation
7-2 C44966AB
Compensation
Creating a Compensation Experiment 7
Creating a Compensation Experiment
Before creating a compensation experiment, you must verify the instrument’s detector
configuration settings (see Verifying, Selecting, Editing, and Creating Detector Configuration in
CHAPTER 6, Data Acquisition and Sample Analysis).
1 Select New Compensation in the File menu or on the start page to create a new compensation
experiment.
NOTE The file name of the newly created compensation experiment has a .xitc suffix.
2 Navigate to the desired file path and select Save. The Compensation Setup window appears.
CAUTION
Risk of erroneous results. Select an unstained tube, according to which the
fluorescence background will be set. If there is not an unstained tube, then
each single color tube must have a negative population.
It is important to specify the appropriate sample type. Otherwise, the
background information could be incorrectly calculated and lead to erroneous
compensation results.
3 Select the channel requiring compensation calculation, and the sample type.
C44966AB 7-3
Compensation
If a negative population is not present in each single color tube, then an unstained control tube
is recommended.
NOTE The default selection is Area. The unstained negative control tube can be selected if needed.
NOTE Label and lot number information can be retained in the Compensation Library to facilitate future
compensation calculations.
7-4 C44966AB
Compensation
4 Select OK.
After confirmation, the software automatically generates the following compensation
experiment.
NOTE Select Area to calculate compensation based on the Area measured. Alternatively, select Height
to calculate compensation based on the Height measured.
Preparing the Compensation Sample

To perform a compensation experiment, prepare:
• Single positive control tube for each color

• Negative control tube (optional)
NOTE A negative control tube is required if a single positive control tube does not contain a negative
population.
For the negative control sample and single positive control sample, you can use blood, cells, or
dedicated compensation beads such as VersaComp Antibody Capture Beads. For details, refer to the
appropriate reagent instructions for use. The negative control tube is used to determine the
autofluorescence of the sample.
C44966AB 7-5
Compensation
Using Control Samples to Generate the Compensation Matrix

Defining the Negative Population Using Unstained Samples
1 Confirm that the instrument has been initialized. Refer to Initializing the Instrument in
CHAPTER 4, Daily Startup.
CAUTION
Risk of erroneous results. Calculations based on excessively small volumes of
sampled data can be inaccurate. Ensure that more than 1,000 positive events and
more than 1,000 negative events are sampled. If the ratio of positive cells is
comparatively low, increase the number of acquisition events to a suitable
amount.
2 Import the gain setting and apply the setting to all tubes. Refer to Adjusting the Gain in
CHAPTER 6, Data Acquisition and Sample Analysis. Use the pan tool to adjust the axis scale so
that the sample signal appears in a suitable position. Adjust the gate so that it encloses the
target cell population (see Creating Plots and Gates in CHAPTER 6, Data Acquisition and Sample
Analysis).
3 Place the negative control tube in the sample tube holder.
4 Select the unstained tube.
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Compensation
6 Set an appropriate number of cells to save in Events to Record located on the left side of the
screen.
Number displayed: 1-500,000
Number recorded: 1-100,000,000
Recording time: ≤1,800 seconds
7 Select Record to save the data.
Running the Single Positive Control Samples
1 Place the single positive tube in sample loading position.
2 Select the appropriate, corresponding tube.
C44966AB 7-7
Compensation
CAUTION
Risk of erroneous results. Calculations based on excessively small volumes of
sampled data can be inaccurate. Ensure that more than 1,000 positive events and
more than 1,000 negative events are sampled. If the ratio of positive cells is
comparatively low, increase the number of acquisition events to a suitable
amount.
4 Move the gate in the FSC/SSC plot so that it encloses the desired population. Move the positive
gate in the plot so that it encloses the positive population. If necessary, move the positive gate
so that it encloses the positive population.
NOTE Figure 7.3 shows an example of selecting the positive population when the negative population
is defined by the unstained sample.
Figure 7.3 Positive Population Selected from the Single-Stained Sample
1. Positive population
7-8 C44966AB
Compensation
NOTE Figure 7.4 shows an example of selecting both the positive and negative populations without an
unstained sample.
Figure 7.4 Positive and Negative Populations Without an Unstained Sample
1. Negative population
2. Positive population
5 Select Record.
6 Repeat steps 1-5 to acquire data from subsequent single positive sample tubes.
CAUTION
Risk of erroneous results. While the software automatically adjusts the
compensation calculation according to gain, excessive adjustment of the
fluorescence gain could lead to inaccurate results.
7 If necessary, adjust gain while acquiring data from single positive sample tubes. Refer to
Adjusting the Gain in CHAPTER 6, Data Acquisition and Sample Analysis.
Calculating Compensation Values
1 Check all acquired sample tubes and confirm that the gating is appropriate.
C44966AB 7-9
Compensation
2 Select or select Compensation Calculation in the Compensation menu to calculate the

compensation values.
The Compensation Matrix window appears, displaying the calculated compensation values.
NOTE The primary fluorescence channels are listed in columns; the secondary fluorescence channels
are listed in rows.
NOTE In the Compensation Matrix window:

• The Use checkbox applies the compensation to the selected sample.
• The Show Autofluorescence checkbox displays the vectors for the autofluorescence.
Numerical value range: -10,000 to 10,000
3 Select Save As to export the compensation matrix as a .comp file and specify where to save it.
NOTE The compensation matrix can also be imported for use in other experiments.
4 Select Save To Compensation Library to save the single color compensation values in the
compensation library.
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Compensation
Creating the Compensation Matrix from Previously Acquired Data 7
5 Specify the key words and select OK.
NOTE The settings stored in the compensation library are specific to the detector configuration. The
compensation library can only be applied when the detector configurations are the same.
At any time, saved compensation experiments can be reopened and the compensation values
recalculated.
6 Select Close.
Creating the Compensation Matrix from Previously Acquired Data
The software supports importing single color data acquired from other experiments into a
compensation experiment to perform compensation calculations. The data to be imported must
match the active detector configuration at the time that the compensation experiment was created.
Otherwise, the data cannot be imported. It is important to ensure that imported data comes from
the same instrument and uses the same configuration and channels. Data originating from a
different instrument will cause erroneous calculations.
1 Select New Compensation from the File menu or the start page.
2 To create a compensation experiment, select the required channels. Refer to Setting the
Channel and Label in CHAPTER 6, Data Acquisition and Sample Analysis.
C44966AB 7-11
Compensation
Adjusting Compensation
3 Right-click on the appropriate test tube and select Import FCS File. Locate the corresponding
data file and import the file. Only files compatible with the detector configuration are
supported by the software for importing.
in front of a test tube indicates that the corresponding data have been imported.
4 After importing the data, adjust the gates to properly identify the positive population and the
negative population for each single-color sample.
5 Calculate the compensation values and export them. Refer to Calculating Compensation Values.
Manually Adjusting Compensation

The compensation can be manually adjusted in an experiment in two ways:
• Select the bivariate plot which the compensation needs to be adjusted. Select from the
graphic control area, then click and drag the mouse pointer up and down or left and right inside
the plot to adjust compensation.
• Select Compensation Matrix in the Setting menu to open the compensation matrix. Adjust the
compensation value between the primary channel and the secondary channel.
7-12 C44966AB
Compensation
Adjusting Compensation 7
Importing and Exporting Compensation
Importing Compensation Settings from Compensation Matrix Files
1 Select the desired sample tube for importing compensation values.
2 Select Compensation Matrix in the Setting menu.
3 Select Import and locate the path where compensation matrix files are saved. Select the
corresponding compensation matrix file (.comp) to import the compensation values.
You can also select Import from Library to import compensation values from the compensation
library. The Import from Compensation Library window appears. Refer to Importing
Compensation Settings from the Compensation Library.
Both methods allow you to import the compensation values with or without the adjustment
based on the gain settings.
4 After opening the desired compensation file, the Import Compensation window appears. Select
one of the following:
• Import compensation matrix and transform it with current gains.
• Import compensation matrix.
• Import compensation matrix and gain.
NOTE
• If the tube does not have any data when importing compensation values calculated from other
instrument settings, the software prompts you to select whether the gain settings must be
imported as well. Select Yes to import fluorescence channel gains settings along with the rest
of the data. Select No to allow the CytExpert for DxFLEX software to adjust the compensation
matrix values based on the current gain settings.
• If the tube does have data when importing compensation values from other instrument settings,
the software prompts you to select whether the compensation values are adjusted based on the
current gain settings.
• It is important to note that automatic adjustments to compensation values calculated from
other instrument gain settings could result in incorrect compensation. Always review the data
after importing compensation values to ensure the sample is compensated properly.
C44966AB 7-13
Compensation
5 Select OK.
6 If necessary, select Apply to to apply the compensation values to the selected test tubes.
7 Select Close.
Importing Compensation Settings from the Compensation Library

You can choose which single color data to include from the compensation library. Only single color
data in the compensation library from the same detector configuration can be imported into the
compensation matrix.
NOTE Files available in the compensation library are configuration-specific. The compensation library only
displays the files created under the current default configuration.
1 Select Import From Compensation Library to select which compensation values to import from
the compensation library.
7-14 C44966AB
Compensation
2 In the Keywords column, the corresponding compensation values can be selected for each
channel. The compensation values of the same keyword can also be selected using the
drop-down menus in the Keywords column.
3 Select OK to import the compensation values.
Exporting Compensation Settings
1 Select the desired sample tube to export.
2 Select Compensation Matrix in the Setting menu.
C44966AB 7-15
Compensation
3 Select Export to specify a path and filename for the compensation file you are saving.
4 Select Save.
NOTE The generated file ends in .comp.
7-16 C44966AB
Compensation
Managing the Compensation Library

Compensation values can be managed in the Compensation Library.
1 Select Compensation Library from the Settings menu. The Compensation Library window
appears.
NOTE The Compensation Library is arranged by fluorescence detection channels.
Tag: String length may not exceed 16
Batch number length: String length may not exceed 16
2 Select the desired single color sample. The compensation information appears on the right side
of the window.
NOTE Existing compensation values (height and area) can be modified by double-clicking the
appropriate column in the Compensation Library window.
3 Enter the Label and Lot No. for the specified single color sample.
4 Select OK.
C44966AB 7-17
Compensation
Adding Channels for Compensation

Channels requiring compensation calculations that have not been previously acquired can be added
to the compensation experiment by acquiring the necessary positive tubes.
1 In the compensation experiment, select in the compensation controls, or select

Compensation Setup in the Compensation menu. The Compensation Setup window appears.
2 Select the channel that needs to be added and select OK.

The software automatically adds a new single positive tube to the compensation experiment. It
also adds a plot with appropriate parameters in the negative control tube.
NOTE It is important to ensure that the data for the previously acquired negative control now includes
the data of the newly added channel and that the settings are correct. Otherwise, you must
reacquire the negative control tube and adjust the gain.
3 Repeat 1-2 to detect and acquire newly added single positive sample data.
4 Repeat Calculating Compensation Values to recalculate and export the compensation results.
7-18 C44966AB
CHAPTER 8
Data Review
Overview
This chapter discusses how to use the Analysis screen to analyze data. Data can be analyzed using
any computer equipped with the CytExpert for DxFLEX software. No online connection is required.
Workflow:
Import experiment Plot and configure Analyze Review

Ô Ô Ô Ô Export results
or data statistics Data Data
• Copying Experiments and Importing Data

• Setting the Plots and Statistics
• Calculating Sample Injection Volume and Concentration
• Adjusting Compensation Settings
• Reports
• Analyzing Samples and Tubes
• Reviewing Samples and Tubes
• Exporting Results
Copying Experiments and Importing Data
Copying a Previously Acquired Experiment

Experiments acquired by other DxFLEX instruments using CytExpert for DxFLEX software can be
imported to your computer for analysis, provided your computer also uses CytExpert for DxFLEX
software.
Select Open Experiment from the Start page or select Open Experiment in the File menu to open the
copied experiment. Then, select Save As.
NOTE The .xit and data folder must be stored in the same path.
Importing Previously Acquired Data

The CytExpert for DxFLEX software can import and analyze compatible FCS data files acquired by
other DxFLEX flow cytometers.
C44966AB 8-1
Data Review
Copying Experiments and Importing Data
1 Create a new experiment or open a saved experiment. Refer to Creating Experiments in

CHAPTER 6, Data Acquisition and Sample Analysis.
2 In the new or opened experiment, select Import FCS File in the File menu to import the data
files.
Imported data files appear in the Tube screen.

The symbol in front of each data tube indicates that the data tube is an imported data file.
Imported data files are copied and saved in the folder where the current experiment data files
are saved.
8-2 C44966AB
Data Review
Setting the Plots and Statistics 8
Setting the Plots and Statistics
Opening the Analysis Screen
1 Select on the left to enter the Analysis screen.
2 Copy plots obtained during data acquisition.
a. If you need original plots used during data acquisition, select to access the Acquisition
screen.
b. Select the appropriate plots.
c. Right-click the selected plots and select Copy from the drop-down menu or press Ctrl+C to
copy.
d. Select to return to the Analysis screen.

e. Select the required test tube from the tube list on the left side of the screen.
f. Right-click the plot area and select Paste from the drop-down menu or press Ctrl+V to paste
the plot.
NOTE Pasted plots include all gates, but the gate names are reassigned.
C44966AB 8-3
Data Review
3 New plots can be created according to need. After selecting the test tubes requiring analysis,
use the plotting control buttons at the top of the screen to create a new plot.
NOTE Each graph in the Analysis screen may correspond to different data. Pay special attention to each
plot’s heading to avoid mistakes during analysis.
4 Use the sample selection controls in the graphics controls toolbar at the top of the page (see
Figure 2.1) to change the data displayed in a plot.
Standard Experiment
Panel Experiment
a. Select the plot requiring a change to the data displayed. By pressing and holding the Ctrl
key while selecting plots, you can select several plots at one time.
b. Select one of the two triangular sample selection buttons ( or ) to choose between
the previous sample and the next sample, or select to specify which data to display.
8-4 C44966AB
Data Review
Setting the Plots and Statistics 8
Creating Multi-Data Histograms and Dot Plot Overlays

The CytExpert for DxFLEX software supports histogram and dot plot data overlay functionality,
allowing you to combine data from differing sources onto the same histogram or dot plot.
1 Select Histogram Overlay under the histogram icon drop-down list to create a new multi-data
histogram.
Or
Select Dot Plot Overlay under the dot plot icon drop-down list to create a new dot plot overlay.
C44966AB 8-5
Data Review
2 Select to select samples for overlay display. Or, drag and drop samples from the tube list on
the left into the histogram or dot plot overlay. The software automatically assigns different
colors to different data.
NOTE In a panel experiment, the sample name displays as [Sample ID] followed by the tube name. For
example [Sample1] Tube1.
To remove a sample, select and uncheck the sample. Or, right-click the color legend and
select Remove [sample name] or Remove All Sample. The corresponding data will no longer
appear on the graph.
3 To change the color selections, right-click on the sample name in the legend located on the
right side of the plot and select Color from the drop-down menu. A color pallet appears.
For configuring gates and generating statistics, refer to CHAPTER 6, Data Acquisition and Sample
Analysis.
8-6 C44966AB
Data Review
Calculating Sample Injection Volume and Concentration 8
Calculating Sample Injection Volume and Concentration
The DxFLEX flow cytometer supports the calculation of the sample concentration based on the
volume consumed and/or based on the known concentration of reference beads.
NOTE If necessary, calibrate the sample uptake rate (see Calibrating the Sample Flow Rate in CHAPTER 12,
Replacement/Adjustment Procedures) prior to collecting data for volumetric analysis:
• Select the cells/μL(V) check box to calculate concentration directly.
NOTE The direct calculation of concentration can be affected by several conditions such as, the
sample’s viscosity and sample mixing. Uncalibrated sample volume uptake rates may lead to
erroneous results.
• If using reference beads to calculate the concentration, select the cells/μL(B) check box and select
the gated Beads Population. Enter the total number of reference beads as the Beads Count, as well
as the sample volume in total. The software automatically calculates the original sample
concentration based on the input values. (You can also enter the reference bead concentration
directly in the beads count field and set the sample volume as 1.)
To obtain accurate calculations, throughout the data acquisition process, ensure that:
• The sample concentration is 2 x 104-107 units/mL.

• Samples are thoroughly mixed before loading and that they exhibit no apparent subsidence
throughout the testing process.
• The detection rate is maintained at less than 10,000 events/second throughout the sampling
process. When the detection rate does not exceed the stated event rate. Running medium to
high acquisition speeds are considered more accurate.
• A constant sampling rate is maintained when recording data.
• You acquire at least 10 μL of sampling volume.
C44966AB 8-7
Data Review
Adjusting Compensation Settings
In the Statistics Setting screen, select Volume and the concentration item to see the corresponding
information in the statistical table.
NOTE While collecting samples, instantaneous data calculation can appear inaccurate. Regard the
calculation as accurate only after data acquisition has been completed.
Number of microspheres: 1-10,000,000, Sample volume: 1-100,000 μL
Adjusting Compensation Settings
Data compensation can be carried out at any time. You can select the desired tube in the tube list
on the left side of the screen and select in the compensation controls, or select Compensation
Setup in the Compensation menu. Refer to Adjusting Compensation in CHAPTER 7, Compensation,
for detailed instructions on adjusting compensation settings.
8-8 C44966AB
Data Review
Reports 8
Reports
Creating a Report
To create a new report, select . A new report tab appears.
NOTE A maximum of 36 reports can be created for each sample.
Copying, Deleting, or Renaming a Report

To copy, delete, or rename a report, right-click the desired report tab and select the desired option
from the menu.
Setting a Report as Default

To set a report as the default report, right-click the desired report tab and select Set as Default. A
checkmark appears in the report tab to denote which report is marked as the default report.
Editing the Report Master Page
Select to switch to Master Page mode. The following message appears at the top of the screen:
Elements added in Master Page mode cannot be edited outside of Master Page mode. The text,
image, property, and page number tools are available in master page mode. Refer to Designing a
Report.
C44966AB 8-9
Data Review
Reports
Designing a Report
Select the desired Report tab. The report displays in the plot area.
NOTE The report toolbar is different for standard experiment and panel experiments. Refer to Figure 8.1 and
Figure 8.2.
Figure 8.1 Report Toolbar - Standard Experiment
1. Test Result 4. Property

2. Text 5. Page Number
3. Image
Figure 8.2 Report Toolbar - Panel Experiment
1. Histogram Overlay 5. Image

2. Dot Plot Overlay 6. Property
3. Test Result 7. Page Number
4. Text
8-10 C44966AB
Data Review
Reports 8
Histogram Overlay
Select from the report toolbar to add a histogram overlay to a report.
Right-click on the histogram image and select Select Tubes to choose which tubes to display in the
overlay.
Dot Plot Overlay
Select from the report toolbar to add a dot plot overlay to a report.
Right-click on the dot plot overlay image and select Select Tubes to choose which tubes to display
in the overlay.
C44966AB 8-11
Data Review
Reports
Test Result
Select from the report toolbar to add a test result element to a report.
To automatically fit the test result frame to the data, right-click the test result element in a report
and select Auto Adjust Height. Refer to Figure 8.3.
Figure 8.3 Test Result Element (Right-Click Menu Shown)
To export test results, right-click the test result element in a report and select the applicable export
option. Refer to Figure 8.3.
To format text in the test result frame, right-click the test result element in a report and select
Property. The Property window appears. Refer to Figure 8.3 and Figure 8.4.
Figure 8.4 Test Result Property Window
To copy or delete a test result element, right-click the test result element in a report and select Copy
or Delete. Refer to Figure 8.3.
8-12 C44966AB
Data Review
Reports 8
Managing the Statistics Settings Displayed in the Test Result Element
1 Right-click the test result element and select Setting. The Statistic Settings window appears.
2 Select Add.
NOTE To delete a statistic setting, select Delete.
C44966AB 8-13
Data Review
Reports
3 Enter a name for the new statistic setting in the name column.
4 Select the empty expression cell then select . The Expression window appears.
5 Enter the desired expression for the selected parameter then select OK.
8-14 C44966AB
Data Review
Reports 8
6 Select the empty reference range cell then select . The Reference Range Setting window
appears.
7 Select one of the range expressions then enter the reference range.
8 Select OK.
9 Enter the unit of measure in the unit cell.
10 Select OK. The new statistic setting is added to the test result window.
Exporting the Test Result Element as a CSV File
1 Right-click the test result element from the report window then select Export to CSV File to
export statistic results or Export Samples to CSV File to export the full report.
C44966AB 8-15
Data Review
Reports
2 Browse to the desired file path then select OK.
Text
IMPORTANT If desired, this element can be added in Master Page mode. Refer to Editing the Report Master
Page.
Select from the report toolbar to add a text box to a report.
To edit the text box, double click the text box.
You can highlight the text then right-click on the highlighted text to undo changes, cut, copy, paste,
delete, or select all.
To change text properties, right-click on the text box and select Property. The Text Property
window appears. Make any desired changes then select OK.
Image
Page.
Select from the report toolbar then browse for the desired image and select OK. The image
displays in the report.
8-16 C44966AB
Data Review
Reports 8
To change an image once it has already been added to the report, right-click the existing image and
select Select Image and browse for a new image.
You can also right-click the image and choose to Lock original aspect ratio, Show Boarder, Copy,
Delete, or Export to Clipboard the image.
Property
Page.
Select the dropdown menu next to the property button on the report toolbar then select a property
element to add to a report.
Standard Experiment Property Options
C44966AB 8-17
Data Review
Reports
Panel Experiment Property Options
Page Number
IMPORTANT This element is only selectable in Master Page mode. Refer to Editing the Report Master Page.
Select from the report toolbar to add a page number element to a report.
8-18 C44966AB
Data Review
Reports 8
Adding and Deleting Pages from a Report
To add a new page to an existing report, select .
To delete an existing page from an existing report, select .
Zooming In and Out of a Report

Use the zoom in/out slider bar located on the bottom, right corner of the report view to zoom in
and out of a report.
NOTE The zoom range is 50% to 300%.
Linking a Plot from a Worksheet to a Report
1 Create a report. Refer to Creating a Report.
2 Select the worksheet containing the desired plot.
NOTE You can select multiple plots to link to the report.
C44966AB 8-19
Data Review
Reports
3 Right-click the desired plot and select Link to Report. The Select Report window appears.
8-20 C44966AB
Data Review
Analyzing Samples and Tubes 8
4 Select which report to link the plot to then select OK. The linked plot appears on the selected
report.
Report Output
Reports can either be printed or exported as a PDF or CSV file via the output icons on the report
toolbar. Refer to Figure 8.5.
Figure 8.5 Output Options on Report Toolbar
Analyzing Samples and Tubes
The Analyze function can be used to analyze a sample as a whole in a panel experiment or an
individual tube in a standard experiment. The Analyze function is available only after the
acquisition is performed.
Standard Experiment
C44966AB 8-21
Data Review
Panel Experiment
IMPORTANT The set analysis time function cannot be performed if:

• The sample or tube is not acquired.
• The sample or tube is damaged.
• There is no tube associated with a sample.
IMPORTANT The tube property can not be changed after the sample or tube is analyzed.
• Ensure a report is created before you analyze or batch analyze sample(s) or tube(s). Refer to Reports
to create a report and design a report.
• If you want to change the settings after an analysis, select Cancel Analysis or Cancel Batch
Analyses to unlock the settings. The analysis information will be cleared. Refer to Cancelling an
Analysis or Canceling Batch Analyses.
Analyzing a Single Sample or Tube
1 Select a sample or tube.
2 Select Analyze from the Analyze and Review dropdown button .
8-22 C44966AB
Data Review
3 The Confirm window appears. Select Yes.
A plot icon appears next to the samples or tubes that have been analyzed.
The analysis information displays in the report if the property elements Analyzed by and
Analysis time have been added to the report.
Cancelling an Analysis
To cancel an analysis, select a sample or tube that has been analyzed then select Cancel Analysis
from the Analyze and Review dropdown button .
The Confirm window appears. Select Yes.
C44966AB 8-23
Data Review
The plot icon disappears from the sample or tube and the analysis information is removed from the
report.
Batch Analyze
The Batch Analyze function can be used to analyze multiple samples as a whole in a panel
experiment or multiple tubes in a standard experiment. The Batch Analyze function is available
only after the acquisition is performed.
1 Select Batch Analyze from the Analyze and Review dropdown button .
8-24 C44966AB
Data Review
The Batch Analyze window appears.
NOTE The available samples or tubes may change depending on the to-be analyzed samples or tubes.
The samples or tubes which are to-be acquired or have been analyzed do not display on the window.
The related message appears below the grid and can be closed.
2 Select the samples or tubes you need to set the analysis time for then select OK.
3 The Confirm window appears. Select Yes.
A plot icon appears next to the samples or tubes that have been analyzed. The analysis
information displays in the report if the property elements Analyzed by and Analysis time have
been added to the report.
C44966AB 8-25
Data Review
Canceling Batch Analyses

To cancel analysis for multiple samples or tubes, select Cancel Batch Analyses from the Analyze and
Review dropdown button . The Cancel Batch Analyses window appears.
NOTE The available samples or tubes may change depending on the samples or tubes which have been
analyzed. The samples or tubes which are to-be acquired, to-be analyzed, or have been reviewed do not
display on the window. The related message appears below the grid and can be closed.
8-26 C44966AB
Data Review
Reviewing Samples and Tubes 8
The plot icon disappears from the samples or tubes. The analysis information is removed from the
report.
Reviewing Samples and Tubes
The review function can be used to review a sample as a whole in a panel experiment or an
individual tube in a standard experiment. The review function is available only after the analysis is
performed.
IMPORTANT The review function cannot be performed if:
• The sample or tube has not been acquired, nor analyzed.
• The sample or tube is damaged.
• There is no tube associated with a sample.
Standard Experiment
C44966AB 8-27
Data Review
Panel Experiment
NOTE Individual tubes can only be reviewed within a standard experiment. In a panel experiment, the review
function is used to review an entire sample. Individual tubes cannot be reviewed in a panel experiment.
IMPORTANT The tube property can not be changed after the sample or tube is reviewed.
• If you want to change the settings after a review, select Cancel Review and then Cancel Analysis
to clear the review/analysis information and unlock the settings.
Reviewing a Single Sample or Tube
1 Select a sample or tube.
2 Select Review from the Analyze and Review dropdown button.
8-28 C44966AB
Data Review
The Review window appears.
3 Optional: Enter any review comments.
4 Select OK. The Confirm window appears.
Select Yes. A checkmark appears next to a sample or tube indicating that the sample or tube has
been reviewed. The reviewer's name and the review time displays in the report.
C44966AB 8-29
Data Review
Canceling a Review
To cancel a review, select a sample or tube that has been reviewed then select Cancel Review from
the Analyze and Review dropdown button.
The checkmark turns back to the plot icon.
Batch Review
The batch review function can be used to review multiple samples as a whole in a panel experiment
or multiple tubes in a standard experiment.
8-30 C44966AB
Data Review
1 Select Batch Review from the Analyze and Review dropdown button.
The Batch Review window appears.
C44966AB 8-31
Data Review
2 Select which samples or tubes to review.
analyzed. The samples or tubes which are to-be acquired, to-be analyzed, or have been reviewed do
not display on the window. The related message appears below the grid and can be closed.
3 Optional: Add review comments in the comment section of the Batch Review window.
Select Yes. A checkmark appears next to a sample or tube indicating that the samples or tubes
have been reviewed. The review information displays in the report.
8-32 C44966AB
Data Review
Canceling Batch Reviews
1 Select Cancel Batch Review from the Analyze and Review dropdown button. The Cancel Batch
Review window appears.
2 Select which samples or tubes to cancel the review from.
reviewed. The samples or tubes which are to-be acquired or to-be analyzed do not display on the
window. The related message appears below the grid and can be closed.
C44966AB 8-33
Data Review
Exporting Results
Select Yes. The review checkmark turns back to the plot icon.
The review information is removed from the report.
Exporting Results
Refer to CHAPTER 6, Data Acquisition and Sample Analysis.
8-34 C44966AB
CHAPTER 9
Daily Shutdown
Overview
This chapter provides procedures for shutting down the DxFLEX instrument.
Workflow:
Prepare the cleaning solution Ô Clean the instrument Ô Turn the instrument off
• Preparing the Cleaning Solution

• Shutting Down the Instrument
Preparing the Cleaning Solution
Required materials
Materials to prepare:
• 12 x 75 mm sample loading tube or a well plate
• FlowClean
• Deionized water
• Bleach (if needed for spills)
Shutting Down the Instrument
1 Run Daily Clean to clean the sample line. Refer to Daily Clean in CHAPTER 11, Cleaning
Procedures.
2 If necessary, empty all waste liquid from the waste container. Refer to Emptying the Waste
Container in CHAPTER 12, Replacement/Adjustment Procedures.
3 Remove the sample tube from the instrument and store according to your laboratory
procedures.
C44966AB 9-1
Daily Shutdown
Shutting Down the Instrument
4 Select Standby.
5 Exit the software.
6 Optional: Turn the computer off.
7 Optional: Turn the Cytometer's main power switch off.
8 If there are any spills, clean the sample station. Refer to Cleaning the Sample Station or
Autoloader in CHAPTER 11, Cleaning Procedures.
9-2 C44966AB
CHAPTER 10
Troubleshooting
Overview
IMPORTANT In addition to the information stated, never disassemble the instrument or have it repaired by
unauthorized personnel. Beckman Coulter bears no responsibility for any problems arising from the
unauthorized repair of the instrument.
This chapter introduces solutions to common problems. If there is a problem, follow the
information in this chapter to carry out self-inspection. If the problem cannot be resolved, contact
us.
• Precautions/Hazards
• Hazard Labels and Locations
• Disposal of Electrical Instrumentation
• RoHS Notice
• Disposal Precaution
• Troubleshooting Table
• Backup/Restore
• Viewing Cytometer Information
Precautions/Hazards
Laser Related Hazards

Beckman Coulter design and manufacture of the instrument complies with the requirements
governing the use and application of a laser specified in regulatory documents issued by the:
• U.S. Department of Health and Human Services

• Center for Devices and Radiological Health (CDRH)
• International Electrotechnical Commission (IEC)
In compliance with these regulatory documents, every measure has been taken to ensure the health
and safety of users and laboratory personnel from the possible dangers of laser use.
Use the instrument according to the information in the manuals.
Use controls or adjustments or performance of procedures other than those specified herein might
result in hazardous radiation exposure.
C44966AB 10-1
Troubleshooting
Precautions/Hazards
To ensure your safety, the Cytometer lasers are covered with protective shields. Do not remove
these shields.
No user-serviceable assemblies are accessible. Do not attempt to remove the laser or open it. The
instrument has components that are dangerous to the operator. If any attempt has been made to
defeat a safety feature, or if the instrument fails to perform as described in its manuals, disconnect
the power and contact us.
Laser Beam Hazards

The flow cytometer can contain up to 3 solid-state diode lasers that are capable of producing laser
light at the following levels:
• 405-nm, 80-mW solid-state diode laser

A laser beam is a unique light source that shows characteristics different from conventional light
sources. The safe use of the laser depends upon familiarity with the instrument and the properties
of coherent, intense beams of light.
WARNING
Risk of personal injury. The laser beam can cause eye damage if viewed either
directly or indirectly from reflective surfaces (such as a mirror or shiny metallic
surfaces). To prevent eye damage, avoid direct exposure to the laser beam. Do not
view it directly or with optical instruments.
Indirect contact with the laser beam from reflective surfaces (such as jewelry or a screwdriver) is
called specular reflection and might also cause damage.
For these reasons, it is important to:

• Limit access to the Cytometer to trained and experienced personnel.
• Never attempt to remove a shield housing a laser.
• Never remove a warning label.
• Contact us if a label is missing or unclear.
Laser Warning Labels
WARNING
Risk of personal injury from radiation exposure. Never remove the shield
surrounding a laser. Never remove covers.
CDRH-approved and IEC compliant labels are also placed near or on those covers that when
removed might expose laser radiation. If necessary, a cover with a CDRH-approved or IEC compliant
label must be removed by a qualified Beckman Coulter Representative only.
10-2 C44966AB
Troubleshooting
Precautions/Hazards 10
Refer to the following figures for the locations of the CDRH-approved and IEC compliant labels:
See Figure 10.1 and Figure 10.2 for the Laser Warning Label on the Cytometer optical bench.
See Figure 10.3 and Figure 10.4 for the Laser Warning Label on the optical bench (located Inside
the Cytometer).
See Figure 10.5 for the Laser Warning Label located inside the DxFLEX Autoloader.
See Figure 10.6 and Figure 10.7 for the Laser Warning Labels located on the back of the
Cytometer.
The laser product is classified as CLASS 1 when all protective measures are in place. This product
complies with 21 CFR Parts 1040.10 and 1040.11 as well as EN60825-1. See Figure 10.1.
Figure 10.1 Laser Warning Label on the Laser Optical Bench [DxFLEX Without Autoloader]
CAUTION
CLASS 3B LASER RADIATION WHEN OPEN AND
INTERLOCKS DEFEATED. AVOID EXPOSURE TO BEAM
PRÉCAUTION
LASER CLASSE 3B RADIATION LASER QUAND
L ' INSTRUMENT EXT OUVERT ET NON VERROUILLE
ÉVITER TOUTE EXPOSITION AU FAISCEAU
LASER RADIATION
CLASS 3B LASER PRODUCT
CAUTION
PRÉCAUTION
Figure 10.2 Laser Warning Label on the Laser Optical Bench [DxFLEX With Autoloader]
CAUTION
PRÉCAUTION
CAUTION
PRÉCAUTION
C44966AB 10-3
Troubleshooting
Precautions/Hazards
Figure 10.3 Laser Warning Label within the Optical Bench (Located Inside the Cytometer) [DxFLEX Without
Autoloader]
LASER RADIATION
LASER RADIATION
CAUTION
OPEN
CAUTION
AND3BINTERLOCKS DEFEATED
CLASS LASER RADIATION WHEN
AVOID
OPEN EXPOSURE
AND TO BEAM
INTERLOCKS DEFEATED
Figure 10.4 Laser Warning Label within the Optical Bench (Located Inside the Cytometer) [DxFLEX With
Autoloader]
LASER RADIATION
LASER RADIATION
CAUTION
10-4 C44966AB
Troubleshooting
Precautions/Hazards 10
Figure 10.5 Laser Label in Autoloader [DxFLEX With Autoloader]
Figure 10.6 Laser Warning Labels on the Cytometer Back Cover [DxFLEX Without Autoloader]
MANUFACTURED

PRODUIT LASER CLASSE 1

MANUFACTURED
REF
SN
EC REP
IVD
C44966AB 10-5
Troubleshooting
Hazard Labels and Locations
Figure 10.7 Laser Warning Labels on the Cytometer Back Cover [DxFLEX With Autoloader]
MANUFACTURED

PRODUIT LASER CLASSE 1

MANUFACTURED
REF
SN
EC REP
IVD
REF
IVD
SN
EC REP
Carefully read the hazard warning labels on the instrument. The hazard labels are located on the
instrument as indicated.
NOTE If a label is missing or unclear, contact us.
Biohazard Label and Location

Figure 10.8 Biohazard Label on the Fluid Containers [DxFLEX Without Autoloader Shown]
䵬⎆Ṋ ᑳ⎆Ṋ
Pএ⾏ᄤ∈
6KHDWK7DQNP',:DWHU
:DVWH
XSSHU XSSHU
⬉य़˖9$&
乥⥛˖+]+]
ORZHU
NOTE The DxFLEX [With Autoloader] uses the same fluidic containers without the fluid container holder.
10-6 C44966AB
Troubleshooting
Hazard Labels and Locations 10
Figure 10.9 Biohazard Label located in the Sample Station and on the Back of the Cytometer [DxFLEX Without
Autoloader]

MANUFACTURED
REF
SN
EC REP
IVD
Figure 10.10 Hazard Label located in the Semi-automatic Sampling Station of the Cytometer [DxFLEX
Without Autoloader]
C44966AB 10-7
Troubleshooting
Figure 10.11 Biohazard Label located in the Sample Station and on the back of the Cytometer [DxFLEX With
Autoloader]

MANUFACTURED
REF
SN
EC REP
IVD
REF
IVD
SN
EC REP
Electrical Shock Hazard Label and Location

Figure 10.12 Electrical Shock Hazard Label by the Power Switch [DxFLEX Without Autoloader]

MANUFACTURED
REF
SN
EC REP
IVD
10-8 C44966AB
Troubleshooting
Hazard Labels and Locations 10
Figure 10.13 Electrical Shock Hazard Label by the Power Switch [DxFLEX With Autoloader]

MANUFACTURED
REF
SN
EC REP
IVD
REF
IVD
SN
EC REP
Caution Labels and Location

Figure 10.14 Caution Labels [DxFLEX Without Autoloader]

MANUFACTURED
REF
SN
EC REP
IVD
Figure 10.15 Caution Labels [DxFLEX With Autoloader]

MANUFACTURED
REF
SN
EC REP
IVD
REF
IVD
SN
EC REP
C44966AB 10-9
Troubleshooting
Disposal of Electrical Instrumentation
Figure 10.16 Hazard Labels in the Autoloader
Disposal of Electrical Instrumentation
It is very important that customers understand and follow

all laws regarding the safe and proper disposal of electrical
instrumentation.
The symbol of a crossed-out wheeled bin on the product is
required in accordance with the Waste Electrical and
Electronic Equipment (WEEE) Directive of the European
Union. The presence of this marking on the product
indicates:
• that the device was put on the European Market after
August 13, 2005 and
• that the device is not to be disposed via the municipal
waste collection system of any member state of the
European Union.
For products under the requirement of WEEE directive,
please contact your dealer or local Beckman Coulter office
for the proper decontamination information and take back
program which will facilitate the proper collection,
treatment, recovery, recycling, and safe disposal of device.
10-10 C44966AB
Troubleshooting
RoHS Notice 10
RoHS Notice
These labels and materials declaration table (the Table of Hazardous Substance's Name and
Concentration) are to meet People's Republic of China Electronic Industry Standard SJ/T11364-2006
"Marking for Control of Pollution Caused by Electronic Information Products" requirements.
RoHS Caution Label
This label indicates that this electronic information product

contains certain toxic or hazardous substances. The center
number is the Environmentally friendly Use Period (EFUP)
date, and indicates the number of calendar years the product
can be in operation. Upon the expiration of the EFUP, the
product must be recycled. The circling arrows indicate the
product is recyclable. The date code on the label or product
indicates the date of manufacture.
Disposal Precaution
WARNING
Risk of biohazardous contamination if you have skin contact with the waste
container, its contents, and its associated tubing. The waste container and its
associated tubing could contain residual biological material and must be handled
with care. Clean up spills immediately. Dispose of the contents of the waste
container in accordance with your local regulations and acceptable laboratory
procedures.
Use universal precautions when working with pathogenic materials. Means must be available to
decontaminate the instrument and to dispose of biohazardous waste.
C44966AB 10-11
Troubleshooting
Troubleshooting Table
Table 10.1 lists problems that you could encounter while running the DxFLEX flow cytometer, the
probable causes of each problem, and the corrective actions. These problems are listed
alphabetically in the Index, under the primary entry “troubleshooting.”
Table 10.1 Troubleshooting
Problem Probable Cause Corrective Action

The Cytometer cannot be • The power cable is not 1. Ensure that the power cable is securely
turned on. securely connected. connected to the back of the Cytometer.
• The fuse is blown. 2. Replace the fuse. Refer to Replacing the
Fuse in CHAPTER 12,
3. If the problem persists, contact us.
The Workstation cannot • The power cable is not 1. Ensure that the power cable is securely
be turned on. securely connected. connected to the back of the Cytometer.
• The Workstation was 2. Unplug the power cable. Wait 10
restarted too fast. seconds, then plug the power cable back
in. Then, restart the computer.
The connection indicator • Data connection error 1. Ensure that the USB data cable is
light in the lower left • The Cytometer is not turned securely connected to the back of the
corner of the software on. Cytometer and the back of the
screen is red and • The Cytometer’s power cable Workstation. Refer to Figure 1.15.
displays Disconnected is disconnected. 2. Restart the software. Restart the
and Error. Workstation. Refer to Initializing the
Instrument in CHAPTER 4, Daily Startup.
3. Turn on the Cytometer using the power
switch on the back of the instrument.
4. Verify that the power cable is securely
connected to the back of the Cytometer.
The alarm does not • The alarm is not working. 1. Ensure that the USB data cable is
sound when the waste • Instrument data connection securely connected to the back of the
container is full or the error. Cytometer and the back of the
sheath fluid container is Workstation. Refer to Figure 1.15.
low and the software 2. Restart the Workstation. Refer to
status display is red. Initializing the Instrument in CHAPTER 4,
Daily Startup.
3. Restart the software.
10-12 C44966AB
Troubleshooting
Troubleshooting Table 10
Table 10.1 Troubleshooting (Continued)

The fluid status • Instrument data connection 1. Ensure that the USB data cable is
information displays red error. securely connected to the back of the
for Sheath and/or Waste • The sensor connection is not Cytometer and the back of the
even though the sheath working properly. Workstation. Refer to Figure 1.15.
fluid container is full and • The sensor does not work 2. Restart the software.
the waste container is properly. 3. Ensure the sheath fluid harness and/or
empty. the waste harness are properly
connected.
4. Verify that the float of the sensor in the
sheath fluid container and/or waste
container moves freely.
5. Replace the sheath fluid harness and/or
the waste harness. Refer to Replacing
the Sheath Fluid Harness and/or Waste
Harness in CHAPTER 12,
The sample tube holder The setting is incorrect. 1. Ensure that the sample injection mode in
cannot move up and the software is in Semi-Automatic
down automatically. Injection mode. Refer to Selecting the
Proper Sample Injection Mode [Without
Autoloader] in CHAPTER 4, Daily Startup.
The sample flow rate is • The sample probe is clogged. 1. Run Prime. Refer to Priming the Flow Cell
unstable. • The sample contains in CHAPTER 12,
aggregates or clumps. Replacement/Adjustment Procedures.
• There are air bubbles in the 2. Run Daily Clean. Refer to Daily Clean in
flow cell. CHAPTER 11, Cleaning Procedures.
• The sample peristaltic pump 3. Clean the sample probe. Refer to
tubing is aged. Cleaning the Sample Probe in
• The sample peristaltic pump CHAPTER 11, Cleaning Procedures.
tubing is not properly 4. Filter the sample using an appropriately
connected. sized mesh aperture filter.
5. Ensure that the sample tubing is properly
connected. Refer to Replacing the
Sample Probe and/or the Sample
Peristaltic Pump Tubing in CHAPTER 12,
6. Replace the sample probe and sample
peristaltic pump tubing. Refer to
Replacing the Sample Probe and/or the
Sample Peristaltic Pump Tubing in
CHAPTER 12, Replacement/Adjustment
Procedures.
C44966AB 10-13
Troubleshooting

The sampling flow rate is • The threshold setting is too 1. Use the manual threshold setting to
too fast. low. increase the threshold. Refer to
• The sample concentration is Adjusting the Threshold in CHAPTER 6,
too high. Data Acquisition and Sample Analysis.
• There are too many sample 2. Dilute the sample and adjust the
fragments. concentration to approximately 106/mL.
• The sheath fluid filter is 3. Filter the sample using an appropriately
clogged. sized mesh aperture filter.
4. Restain the sample.
5. Replace the Sheath Fluid Filter. Refer to
Replacing the Sheath Fluid Filter in
Procedures.
Laser power is low. Communication error 1. Reinitialize. Refer to Initializing the
2. Restart the Cytometer.
Populations are drifting. • Air bubbles are in the flow 1. Run Prime. Refer to Priming the Flow Cell
cell. in CHAPTER 12,
• Air bubbles are in the Replacement/Adjustment Procedures.
system. 2. Ensure that the sheath fluid harness
• The sheath fluid harness float and/or waste harness is not kinked.
is restricted. 3. Ensure that the sheath fluid harness
and/or waste harness is securely
connected.
4. Run Prime. Refer to Priming the Flow Cell
in CHAPTER 12,
sheath fluid container moves freely.
6. Replace the sheath fluid harness. Refer
to Replacing the Sheath Fluid Harness
and/or Waste Harness in CHAPTER 12,
10-14 C44966AB
Troubleshooting

Population amplitude is • Air bubbles are in the flow 1. Run Prime. Refer to Priming the Flow Cell
decreasing and CV cell. in CHAPTER 12,
values are increasing. • The flow cell is dirty. Replacement/Adjustment Procedures.
• The sheath fluid harness float 2. Run the Deep Clean procedure. Refer to
is restricted. Deep Clean Procedure in CHAPTER 11,
Cleaning Procedures.
The laser delay values • Air bubbles are in the flow 1. Run Prime. Refer to Priming the Flow Cell
are out of range. cell. in CHAPTER 12,
• Air bubbles are in the Replacement/Adjustment Procedures.
system. 2. Ensure that the sheath fluid harness
• The sheath fluid harness float and/or waste harness is not kinked.
is restricted. 3. Ensure that the sheath fluid harness
connected.
C44966AB 10-15
Troubleshooting

No data acquisition. • The threshold setting is too 1. Decrease the threshold setting. Refer to
high. Adjusting the Threshold in CHAPTER 6,
• The gain setting is too low. Data Acquisition and Sample Analysis.
• Sheath fluid flow is 2. Increase the gain setting. Refer to
insufficient. Adjusting the Gain in CHAPTER 6, Data
• Laser power is insufficient. Acquisition and Sample Analysis.
• The sample probe is clogged. 3. Ensure that the sheath fluid harness
and/or waste harness is not kinked.
4. Ensure that the sheath fluid harness
connected.
in CHAPTER 12,
6. Reinitialize. Refer to Initializing the
7. Restart the Cytometer.
8. Verify that your sample does not have
excessive debris. If it does:
a. Filter the sample using an
appropriately sized mesh aperture
filter.
b. Restain the sample.
9. Clean the sample probe. Refer to
Cleaning the Sample Probe in
10. Replace the sample probe and the
sample peristaltic pump tubing. Refer to
Procedures.
Data populations are The laser delay setting is 1. Ensure that the laser delay is set
normal on one laser, but incorrect. correctly. Refer to Setting Laser Delay in
too low on another laser. CHAPTER 12, Replacement/Adjustment
Procedures.
10-16 C44966AB
Troubleshooting

Data populations are not • The detector configuration 1. Ensure that the detector configuration is
where they are setting is incorrect. set correctly. Refer to Verifying,
expected. • The optical filter is not placed Selecting, Editing, and Creating Detector
correctly. Configuration in CHAPTER 6, Data
• QC was not completed. Acquisition and Sample Analysis.
• Gain and threshold is not set 2. Ensure that the position of the optical
correctly. filter in the WDM matches the detector
configuration setting. Refer to Verifying,
Selecting, Editing, and Creating Detector
Configuration in CHAPTER 6, Data
3. Ensure that the optical filter is installed
correctly. Refer to Replacing the Optical
Filter in CHAPTER 12,
4. Follow the QC procedure. Refer to
CHAPTER 5, Instrument Quality Control
and Standardization.
5. Review the gain and threshold settings.
Refer to Adjusting the Gain and Adjusting
the Threshold in CHAPTER 6, Data
6. Review the display ranges. Refer to
Creating Plots and Gates in CHAPTER 6,
Data Acquisition and Sample Analysis.
No changes occurred Compensation was applied to Ensure that the adjustment is applied to the
after manually adjusting the wrong channel. correct primary and secondary channels in
compensation settings. the compensation matrix.
Or
Select to modify compensation in the

desired plot.
C44966AB 10-17
Troubleshooting

The calculation of the • Erroneous data acquisition. 1. Ensure that the corresponding negative
automatic compensation • The gate is not set on the control tube and the individual positive
experiment is incorrect. appropriate population. tube acquired are from the same sample
• The events of the acquired type.
cells are too low. 2. Ensure that the single colors collected
• The mean fluorescence of correspond to the correct tube name.
the positive cells is too weak. 3. Ensure that the gate in the FSC/SSC plot
encloses the correct sample population.
4. Ensure that the positive gate in each
tube is correctly placed.
5. Modify the events to record to ensure
that enough events are collected for the
data population.
6. Select samples with a stronger positive
signal as the positive control.
Or
Use dedicated compensation beads such
as VersaComp Antibody Capture Beads
The sample is flowing, • The signal is outside of the
but no signal appears in display range. 1. Use either or to modify the
the plot. display range.
• The parent gate is not
positioned properly and does Or
not contain events. Right-click the plot and select Property.
• The population color setting The Plot Property window appears.
is too light. Select Fit with sample.
• The threshold is too high. 2. Ensure that the parent gate is gated
correctly.
3. Change the display color.
4. Move the sample above the threshold
using one of the following methods:
• Decrease the threshold setting. Refer
to Adjusting the Threshold in
CHAPTER 6, Data Acquisition and
Sample Analysis.
• Increase the gain setting. Refer to
Adjusting the Gain in CHAPTER 6,
Data Acquisition and Sample
Analysis.
10-18 C44966AB
Troubleshooting

The concentration • The sample concentration is 1. Ensure that the pipette used in sample
calculation is incorrect. not within the specified processing is calibrated.
range. 2. Verify that the concentration of the
• The sample settled. sample is between 2x104-107 events/mL.
• The sample flow rate is too 3. Vortex the sample before loading and
fast. verify that the sample is evenly mixed
• The sample volume analyzed before loading the sample.
is too low.
NOTE An excessively long sample
• The cell population is not
loading time leads to sample
detected.
settlement.
4. Ensure that the sample flow rate does

not exceed 10,000 events/second.
5. Adjust the threshold to remove sample
debris.
6. Ensure that the sample volume analyzed
exceeds 10 μL.
7. Ensure that the following are correct:
• Gain. Refer to Adjusting the Gain in
CHAPTER 6, Data Acquisition and
Sample Analysis.
• Threshold. Refer to Adjusting the
Threshold in CHAPTER 6, Data
• Compensation settings. Refer to
CHAPTER 7, Compensation.
8. Ensure that the gating and population
hierarchy are correct.
The sample probe is too The sample probe is not securely 1. Ensure that the sample probe is securely
low. connected. connected to the sample peristaltic
pump tubing. Refer to Replacing the
Sample Probe and/or the Sample
2. Ensure the sample pump cover is
securely fastened.
The wash station drips • The sample probe is not 1. Ensure that the sample probe is securely
during backflush. securely connected. connected to the sample peristaltic
• The wash station height pump tubing. Refer to Replacing the
adjustment is not correct. Sample Probe and/or the Sample
2. Ensure the sample pump cover is
securely fastened.
C44966AB 10-19
Troubleshooting

The mixer is not • Sample mixing is disabled in 1. Ensure sample mixing is enabled in the
functioning. the software. software. Refer to Changing Sample
• The mixer motor is defective. Mixing and Backflush Settings in
Procedures.
Instrument operations • The instrument is in standby 1. Select Initialize.
cannot be performed in mode. 2. Ensure that the power switch to the
the Acquisition screen. • The software is frozen. Cytometer is turned on.
• Data connection error. 3. Restart the software.
4. Restart the Workstation.
5. Ensure that the USB data cable is
securely connected to the back of the
Cytometer and the back of the
Workstation. Refer to Figure 1.15.
Software installation Multiple issues. Contact us.
fails.
QC aborted due to low • The sample probe is clogged. 1. Reload the target value file. Refer to
event rate. • The sample line is clogged. Importing Lot-Specific Target Values in
CHAPTER 5, Instrument Quality Control
and Standardization. Then, rerun QC.
2. Prepare a new sample of the DxFLEX
Daily QC Fluorospheres. Then, rerun QC.
3. Clean the sample probe. Refer to
Cleaning the Sample Probe in
CHAPTER 11, Cleaning Procedures. Then,
rerun QC.
in CHAPTER 12,
Then, rerun QC.
5. Run Daily Clean. Refer to Daily Clean in
rerun QC.
6. Replace the sample probe and sample
peristaltic pump tubing. Refer to
Procedures. Then, rerun QC.
7. Is problem persists, contact us.
10-20 C44966AB
Troubleshooting

QC failed. • The median fluorescence 1. Rerun QC. Refer to CHAPTER 5,
fails to meet the target Instrument Quality Control and
specification. Standardization.
• The QC gain value does not 2. Run Prime. Refer to Priming the Flow Cell
meet the target gain in CHAPTER 12,
specifications. Replacement/Adjustment Procedures.
• The laser delay settings are Then, rerun QC.
too high. 3. Prime the sheath fluid filter with sheath
• rCV fails specifications. fluid as follows, then rerun QC.
a. Remove the vent cap from the
sheath fluid filter.
b. Ensure that the instrument is in
Standby.
c. At the Workstation, select
Cytometer > Prime.
IMPORTANT If the vent cap is not

reinstalled as soon as the sheath
fluid approaches the vent port, the
sheath fluid will overflow.
d. Wait until the sheath fluid

approaches the vent port in the
sheath fluid filter, then immediately
reinstall the vent cap to avoid
overflow.
4. Run Daily Clean. Refer to Daily Clean in
rerun QC.
5. Run Deep Clean. Refer to Deep Clean
Procedure in CHAPTER 11, Cleaning
Procedures. Then, rerun QC.
C44966AB 10-21
Troubleshooting

The acquisition has • The incorrect sample 1. Verify that the sample injection mode is
stopped abnormally. The injection mode is selected. set correctly. Refer to Selecting the
alarm is sounding and • The Autoloader cover is open Proper Sample Injection Mode [with
the indicator light is red. during a sample acquisition. Autoloader] in CHAPTER 4, Daily Startup.
• The Autoloader is unable to 2. Ensure the Autoloader cover is closed.
move. 3. Verify that the Autoloader is clear of any
— A foreign object is in the foreign objects.
Autoloader. 4. Ensure the tube is seated correctly in the
— The tube is not correctly carousel. Refer to Putting Sample Tubes
seated. in a Carousel in CHAPTER 1, System
— The carousel or the plate Overview.
adapter is not correctly 5. Ensure the carousel or the plate adapter
seated. is seated correctly in the carousel. Refer
to Putting Sample Tubes in a Carousel in
• The USB cable is
CHAPTER 1, System Overview.
disconnected.
6. Ensure that the USB cable is securely
• USB communication error
connected.
• The Autoloader is defective.
7. Verify that the tube location is set
• The tube is missing at the correctly. Refer to Creating Experiments
specified location. in CHAPTER 6, Data Acquisition and
• The carousel number does Sample Analysis.
not match with the carousel 8. Verify that the carousel number is set
barcode. correctly to match the barcode on the
• The sample ID does not actual carousel. Refer to Creating
match the tube barcode. Experiments in CHAPTER 6, Data
9. Verify that the sample ID matches the
tube barcode.
10. Restart the workstation.
The plate adapter or Acquisition was aborted before a 1. Close Autoloader cover.
carousel is stuck in the task was complete. 2. Select Cytometer > Sampler Reset.
sampling position.
The barcode cannot be • The barcode label is dirty. 1. Ensure that the barcode label is clean or
read. • Communication error replace the barcode label.
• Barcode reader is defective 2. Restart the workstation.
10-22 C44966AB
Troubleshooting
Backup/Restore 10
Backup/Restore
CAUTION
Risk of data loss. The database backup does not include experiment files. Save a
copy of the experiment files in a secure location separately.
Use the backup function to back up the CytExpert for DxFLEX database. System backup includes
user account information, log information, and software settings.
Backup
1 Select Backup/Restore > Backup. The Backup window appears.
2 Select a backup directory location.
3 Select Next.
C44966AB 10-23
Troubleshooting
Backup/Restore
The system begins backing up the database.
10-24 C44966AB
Troubleshooting
Backup/Restore 10
4 Select Finish.
Use the restore function to restore the CytExpert for DxFLEX database.
Restore
1 Select Backup/Restore > Restore. The Restore window appears.
2 Select Next. The Select the restore point window appears.
3 Select the most recent backup directory.
C44966AB 10-25
Troubleshooting
Backup/Restore
4 Select Next.
The system begins backing up the database.
10-26 C44966AB
Troubleshooting
Viewing Cytometer Information 10
5 Select Finish.
Viewing Cytometer Information
Select Cytometer > Cytometer Information to display the Cytometer Information window. The
Cytometer Information window displays the following information:
• Product
• Serial Number
• DAQ Version
• MCB Version
• AutoLoader Version
Figure 10.17 Cytometer Information Window
C44966AB 10-27
Troubleshooting
Viewing Cytometer Information
10-28 C44966AB
CHAPTER 11
Cleaning Procedures
Overview
This chapter describes how to carry out certain routine and nonscheduled cleaning procedures.
Proper cleaning can help extend the service life of the instrument and ensure experimental
accuracy. When conducting any cleaning, take all necessary biosafety precautions and use proper
personal protective equipment.
• Routine Cleaning
— Daily Clean
— Cleaning the Sample Station or Autoloader
— Deep Clean Procedure
— Cleaning the Sheath Fluid Container
— Cleaning the Waste Container
• Nonscheduled Cleaning
— Preparing the Instrument for Transport or Storage
Routine Cleaning
Daily Clean
Daily Clean should be performed during instrument startup and instrument shutdown to clean the
sample line.
After sampling an excessively large sample or a sample that can easily clog the sample probe, it is
recommended to perform the Daily Clean procedure.
Refer to Daily Clean [With Autoloader - Plate Mode] in APPENDIX C, DxFLEX [With Autoloader -Plate
Mode] for instructions on running Daily Clean procedures in Plate Mode with the DxFLEX
Autoloader.
C44966AB 11-1
Cleaning Procedures
Routine Cleaning
Daily Clean [Without Autoloader]
1 Open the CytExpert for DxFLEX software and confirm that the instrument is connected and that
it has already been initialized. Refer to Opening the Software in CHAPTER 4, Daily Startup.
2 Select Daily Clean in the Cytometer menu.
3 Add 2 mL of FlowClean solution to an unused sample tube.
4 Add 2 mL of DI water to an unused sample tube.
11-2 C44966AB
Cleaning Procedures
Routine Cleaning 11
5 Insert the sample tube with 2 mL of FlowClean solution into the sample holder and select Run.
NOTE The default cleaning time is 3 minutes.
6 Remove the Flow Clean tube.
C44966AB 11-3
Cleaning Procedures
Routine Cleaning
7 Insert the sample tube with 2 mL of DI water into the sample holder and select Run to perform
the second step of the cleaning process.
NOTE The default cleaning time is 5 minutes.
8 After the process has been completed, remove the sample tube and close the Daily Clean
window.
Daily Clean [With Autoloader - Single Tube and Carousel Mode]
11-4 C44966AB
Cleaning Procedures
Routine Cleaning 11
3 Select Daily Clean in the Cytometer menu. The Daily Clean window appears.
C44966AB 11-5
Cleaning Procedures
Routine Cleaning
IMPORTANT You must select at least one cleaning solution well and one water well.
4 Follow the on screen software prompts and select the desired wells for cleaning agent and
deionized water.
The carousel sets the tube location in positions 1 and 2 by default. indicates cleaning agent
and indicates deionized water.
NOTE To deselect tubes, select the desired tube and select Set As Empty.
NOTE To select new cleaning agent tube locations, select the desired tubes and select Cleaning Agent.
To select new deionized water tube locations, select the desired tubes and select Deionized Water.
5 Select Start to start the cleaning procedure.
NOTE Select Stop at any time to stop the daily clean process.
6 When daily clean is complete, select Close.
11-6 C44966AB
Cleaning Procedures
Routine Cleaning 11
Cleaning the Sample Station or Autoloader
Carry out semi-automatic cleaning for the sample injection device or autoloader once a week.
1 Ensure that the system has been shut down properly. Refer to Shutting Down the Instrument
in CHAPTER 9, Daily Shutdown.
WARNING
the chemical.
2 Use a piece of soft cloth with a 10% bleach solution (1 part bleach [5 to 6% sodium
hypochlorite - available chlorine] with 9 parts DI water) to wipe off all surfaces in the sample
station or autoloader including the wash station, while taking all necessary biological safety
precautions.
[Without Autoloader]
C44966AB 11-7
Cleaning Procedures
Routine Cleaning
[With Autoloader]
Cleaning the Sample Probe
When problems such as blockage of the sample probe occur, it is required to replace or clean the
sample probe.
1 Confirm that the instrument is in the standby state or that the power supply is turned off.
WARNING
Risk of biohazardous contamination if you have skin contact with the sample
probe or the sample peristaltic pump tubing. The sample probe and the sample
peristaltic pump tubing might contain residual biological material and must be
handled with care. Clean up spills immediately. Dispose of the sample probe and
the sample peristaltic pump tubing in accordance with your local regulations and
acceptable laboratory procedures.
2 Remove the sample probe. Refer to Replacing the Sample Probe and/or the Sample Peristaltic
Pump Tubing in CHAPTER 12, Replacement/Adjustment Procedures.
3 Put the sample probe into a clean container and soak it in clean water. Use an ultrasonic
cleaning device to clean for 2 minutes.
11-8 C44966AB
Cleaning Procedures
Routine Cleaning 11
4 Reattach the sample probe to the sample peristaltic pump tubing, and confirm that the bead on
the sample probe touches the sleeve on the end of the sample peristaltic pump tubing.
IMPORTANT To ensure that the proper placement of the sample probe, push the sample pump cover up
while installing the thumbscrew.
5 Install the sample pump cover.
6 If ineffective, replace with a new sample probe. Refer to Replacing the Sample Probe and/or the
Sample Peristaltic Pump Tubing in CHAPTER 11, Cleaning Procedures.
Deep Clean Procedure
Carry out a deep clean once a month to clean the instrument flow cell. If the unit will be shut down
and not used for more than 10 days, it is recommended to complete one deep clean before resuming
use.
1 Place the instrument in standby state.
2 Remove the right-side cover. Refer to Right-Side Cover Removal and Reinstallation in
3 Confirm that the Deep Clean solution volume in the bottle located inside the Fluidics module is
sufficient.
To prepare and add more Deep Clean solution, refer to Adding the Deep Clean Solution in
C44966AB 11-9
Cleaning Procedures
Routine Cleaning
4 Select Deep Clean in the Cytometer menu. The software message Are you sure to start deep clean?
appears. Select Yes to start the Deep Clean process in the instrument flow cell.
5 The status bar prompts that a deep clean is under way. Wait for the Deep Clean process to finish.
The following software message appears:
Select OK.
6 Allow the cleaning solution to remain in the flow cell for approximately 30 minutes. If you are
required to postpone the cleaning time, do not exceed 24 hours. During the Deep Clean cycle,
the power supply of the unit can be turned off, but the instrument cannot be initialized.
11-10 C44966AB
Cleaning Procedures
Routine Cleaning 11
7 Select Prime in the Cytometer menu. The software message Are you sure to start Prime? appears.
Select Yes.
8 Run Daily Clean. Refer to Daily Clean.
9 Perform initialization as required (see Initializing the Instrument in CHAPTER 4, Daily Startup)
to carry out the next experiment or to turn off the instrument.
10 Reinstall the right-side cover. Refer to Right-Side Cover Removal and Reinstallation in
Cleaning the Sheath Fluid Container
CAUTION
DxFLEX [With Autoloader]: Risk of instrument damage. The fluid containers
should be located directly behind the Autoloader. Take care not to kink or unplug
the Autoloader USB connector when moving the fluid containers.
Clean the sheath fluid container once a month.
1 Confirm that the instrument is turned off or is in the standby state.
C44966AB 11-11
Cleaning Procedures
Routine Cleaning
2 DxFLEX [Without Autoloader]: Remove the sheath fluid container from the Fluid Container
holder.
3 Remove the sheath fluid harness from the sheath fluid container.
4 Empty the residual sheath fluid from the sheath fluid container.
5 Add about 50 to 100 mL of DxFLEX Sheath Fluid to the sheath fluid container.
6 Insert the sheath fluid harness back into the sheath fluid container and tightly close the sheath
fluid container cap.
7 Swirl the sheath fluid in the sheath fluid container, rinsing all surfaces.
8 Empty the sheath fluid container.
9 Refill the sheath fluid container. Refer to Filling the Sheath Fluid Container in CHAPTER 12,
Cleaning the Waste Container
CAUTION
Clean the waste container once a month.
2 DxFLEX [Without Autoloader]: Remove the waste container from the Fluid Container holder.
11-12 C44966AB
Cleaning Procedures
Routine Cleaning 11
WARNING
with care. Clean up spills immediately.
3 Remove the harness from the waste container.
WARNING
procedures.
4 Empty the waste container.
WARNING
the chemical.
5 Add one liter of sodium hypochlorite solution with 0.5% active chlorine to the waste container.
6 Insert the waste harness back into the waste container and tightly close the waste container
cap.
CAUTION
Risk of damage to the sheath fluid harness and/or waste harness. Do not leave the
sodium hypochlorite solution in the fluid containers longer than 10 minutes.
7 Let stand for 5 to 10 minutes.
C44966AB 11-13
Cleaning Procedures
Nonscheduled Cleaning
WARNING
the chemical.
8 Dispose of the sodium hypochlorite solution in accordance with your local regulations and
9 Use deionized water to rinse the waste container and the waste harness. Ensure that there is no
sodium hypochlorite residue.
10 DxFLEX [Without Autoloader]: Place the waste container back into the Fluid Container holder.
Preparing the Instrument for Transport or Storage
IMPORTANT If you have a DxFLEX equipped with an autoloader that needs to be transported, contact us.
When the instrument is to be transported or is not to be used for 30 days or more, complete the
emptying processes to prevent instrument damage and to reduce the possibility of biological
contamination.
1 Run the Deep Clean procedure. Refer to Deep Clean Procedure.
2 Run the Daily Clean procedure. Refer to Daily Clean.
3 Clean the Sample Station. Refer to Cleaning the Sample Station or Autoloader.
4 Empty the sheath fluid container and the waste container (see Emptying the Waste Container
in CHAPTER 12, Replacement/Adjustment Procedures).
11-14 C44966AB
Cleaning Procedures
Nonscheduled Cleaning 11
WARNING
procedures.
5 Clean the sheath fluid container and the waste container. Refer to Cleaning the Sheath Fluid
Container.
6 Remove the right-side cover (see Right-Side Cover Removal and Reinstallation in CHAPTER 12,
Replacement/Adjustment Procedures).
7 Remove the Deep Clean solution bottle from the bracket, empty the Deep Clean solution bottle,
and rinse with DI water. Then, attach the Deep Clean solution bottle to the bracket.
8 Reinstall the right-side cover (see Right-Side Cover Removal and Reinstallation in CHAPTER 12,
Replacement/Adjustment Procedures).
9 Power down and disconnect all the cables and sheath fluid and waste harnesses.
10 Ensure that the optical filters are seated properly.
11 Ensure that the top cover is tightly closed.
12 If the instrument is to be transported or stored, put the instrument into the Beckman Coulter
packing and comply with the requirements in Instrument Transportation and Storage in
APPENDIX A, Instrument Installation, regarding correct placement during transportation and
storage.
IMPORTANT If you have a DxFLEX equipped with an autoloader that needs to be transported, contact us.
Lifting and Carrying Instructions [Without Autoloader Only]
1 Position a person on the left and right sides of the Cytometer.
C44966AB 11-15
Cleaning Procedures
2 Reach under the base of the Cytometer in the areas indicated by the arrows in the figure below.
3 Gently lift the Cytometer as shown in the figure below.
WARNING
Risk of personal injury. Use caution when lowering the Cytometer to avoid
pinching fingers.
4 Lower the Cytometer to its designated location.
11-16 C44966AB
CHAPTER 12
Replacement/Adjustment Procedures
Overview
This chapter describes how to carry out certain routine and nonscheduled maintenance
procedures. Proper maintenance can help extend the service life of the instrument and ensure
experimental accuracy. When conducting any maintenance work, take all necessary biosafety
precautions.
IMPORTANT Unless otherwise specified, for all replacement parts, use only parts provided by Beckman
Coulter to ensure proper functioning of the instrument. Never disassemble any part of the instrument
without prior authorization. Beckman Coulter assumes no responsibility for any instrument problems
resulting from the use of any part not authorized by Beckman Coulter for use with the instrument.
• Routine Replacement/Adjustment
— Front Cover Removal
— Right-Side Cover Removal and Reinstallation
— Top and Front Cover of DxFLEX Autoloader Removal and Reinstallation
— Filling the Sheath Fluid Container
— Emptying the Waste Container
— Managing the Maintenance Reminder
— Adding the Deep Clean Solution
— Replacing the Sheath Fluid Filter
— Replacing the Sample Probe and/or the Sample Peristaltic Pump Tubing
— Inspecting the Liquid Flow Path for Leaks
— Priming the Flow Cell
— Changing the Event Rate Setting
• Nonscheduled Replacement/Adjustment
— Calibrating the Sample Flow Rate
— Setting Laser Delay
— Replacing the Optical Filter
— Replacing the Fuse
— Replacing the Sheath Fluid Harness and/or Waste Harness
— Changing Sample Mixing and Backflush Settings
— Calibrating the Carrier [DxFLEX With Autoloader]
C44966AB 12-1
Routine Replacement/Adjustment
Front Cover Removal
Removal
WARNING
Risk of personal injury from electric shock caused by contacting exposed
electronic components. Power down the instrument before removing the front
cover of the Cytometer.
1 Exit the system software.
2 Turn off the main power switch on the back of the Cytometer.
3 Open the top cover.
12-2 C44966AB
Routine Replacement/Adjustment 12
4 Remove the two screws securing the front cover and pull the front cover forward.
CLA
OP
EN SS 3B CAUT
AVOIDAND LAS
INT ER
EXPO ION
ERLO RADIA
SURE CKS TIO
TO DEFEAN WH
BEAM TEDEN
1. Securing screws
2. Front cover
5 Lift the front cover up and out of the slots in the frame.
Reinstallation
1 Slide the tabs on the bottom of the front cover into the slots in the bottom of the frame.
2 Push in the latches on the front cover to retract the pins, push the front cover into place, and
release the latches to secure the cover.
3 Close the top cover.
C44966AB 12-3
Right-Side Cover Removal and Reinstallation
Removal
1 Open the top cover.
2 Unfasten the two captive thumbscrew [Without Autoloader] or one captive thumbscrew [With
Autoloader] for the right-side cover.
c b
b c
E TO EN
BEA ED
UR DEF WH
EAT
OS KS N
ION
EXP LOC IATIO
M
ID NTER RAD
UT
CA
AVO I ER
LAS
OP SS 3B
AND
CLA
EN
1. Right-side cover
2. Captive thumbscrews
12-4 C44966AB
3 DxFLEX [With Autoloader]: Pull and rotate the latch located inside the right-side cover into the
horizontal position.
4 Lift the right-side cover up and out of the slots in the frame.
b
b
CLASS
OPEN CAUT
3B LASER
AND INTERLO
AVOID ION
CAUTION
RADIATI
EXPOSU CKS ON WHEN
CLASS
RE TO DEFEAT
OPEN ED3B LASER
BEAM
AND INTERLO RADIATIO
AVOID EXPOSUR N WHEN
CKS DEFEATE
E TO BEAM D
c
c
1. Right-side cover
2. Slots
C44966AB 12-5
Reinstallation
1 Slide the tab on the bottom of the right-side cover into the slots in the bottom of the frame and
push the cover into place.
CLASS
OPEN CAUT
ION
3B LASER
AND INTERLO
AVOID
CAUTION
RADIATIO
EXPOSU CKS
CLASS
N WHEN
RE TO DEFEATE
OPEN D3B LASER
BEAM
AND INTERLOC RADIATIO
AVOID EXPOSUR N WHEN
KS DEFEATE
E TO BEAM D
c c
1. Tabs
2. Slots
2 DxFLEX [With Autoloader]: Pull and rotate the latch located inside the right-side cover into the
vertical position.
12-6 C44966AB
3 Fasten the captive thumbscrew(s) to secure the right-side cover.
c b
b c
E TO EN
BEA ED
UR DEF WH
EAT
OS KS N
ION
EXP LOC IATIO
M
ID NTER RAD
UT
CA
AVO I ER
EN LAS
OP SS 3B
AND
CLA
1. Right-side cover
2. Captive thumbscrews
4 Close the top cover.
Top and Front Cover of DxFLEX Autoloader Removal and Reinstallation

Removal
1 Unfasten the two captive thumbscrews for the top cover of the autoloader.
C44966AB 12-7
2 Lift the top cover of the autoloader off.
3 Remove the front cover of the autoloader.
Reinstallation
1 Install the front cover of the autoloader.
2 Align the tab of the top cover for the autoloader with the slot in the frame.
12-8 C44966AB
3 Fasten the two captive thumbscrews to secure the top cover of the autoloader.
Filling the Sheath Fluid Container
CAUTION
1 If necessary, remove any cardboard cutouts from the new DxFLEX Sheath Fluid cubitainer. If
you do not need a new cubitainer, skip to Step 6.
2 Locate the spigot inside the cardboard cutout.
3 Remove the cap and seal from the new sheath cubitainer. Be sure to completely remove the foil
seal.
4 Screw on the spigot.
5 Unscrew the sheath fluid harness from the sheath fluid container and place in the fluid sensor
holder cutout (refer to Figure 1.7.) to prevent contamination of the sheath fluid harness.
CAUTION
Risk of instrument damage. Remove the sheath fluid container from the Fluid
Container holder and fill away from the instrument to prevent spills that could
damage the instrument circuitry.
6 DxFLEX [Without Autoloader]: Remove the sheath fluid container from the Fluid Container
holder.
7 Hold the Sheath fluid container under the DxFLEX Sheath Fluid cubitainer ensuring the
cubitainer is resting on a stable surface.
C44966AB 12-9
8 Fill the sheath fluid container.
9 Reinstall the sheath fluid harness onto the sheath fluid container.
10 DxFLEX [Without Autoloader]: Place the sheath fluid container back into the Fluid Container
holder.
Emptying the Waste Container
CAUTION
2 Remove the waste harness (see Figure 1.7). The waste harness from the Cytometer is connected
to a 4-L waste container.
3 DxFLEX [Without Autoloader]: Remove the waste container from the Fluid Container holder.
WARNING
associated tubing might contain residual biological material and must be handled
procedures. Use proper personal protective equipment.
4 Empty the waste container. Dispose of the waste in accordance with your local regulations and
12-10 C44966AB
WARNING
the chemical.
5 Add 400 mL of 5 to 6% bleach to the waste container.
6 Reinstall the waste harness in the waste container.
7 DxFLEX [Without Autoloader]: Put the waste container in the Fluid Container holder.
Managing the Maintenance Reminder

The maintenance reminder tracks the last maintenance date and initiates a reminder to complete
maintenance for the following three items:
• Refill Deep Clean solution bottle

• Replace sheath fluid filter
• Replace sample peristaltic pump tubing
When reagents or parts have reached the designated use time limit in either days or number of uses,
the Maintenance Message icon appears

in the right side of the status bar.
1 Select the Maintenance Message icon from the status bar to access the
Maintenance window. The expired item appears with a warning triangle to the left of the
item listed.
Or
C44966AB 12-11
Select Maintenance in the Advanced menu. The Maintenance window appears. The expired
item is displayed with a warning triangle to the left of the item listed.
2 Select the desired item to manage, then choose one of the following:
• To manage refilling the Deep Clean solution bottle, go to Step 3.
• To manage replacing the sheath filter, skip to Step 4.
• To manage replacing the sample peristaltic pump tubing, skip to Step 5.
3 Select Detail. The Refill Deep Clean Solution Bottle window appears.
Select Refill. A pop-up window appears to reset the maintenance date as the current date that
the maintenance was performed.
Explanation of restrictions (if fluidic self-check is enabled):

Usage time limit: 1-180
Refill volume: 20-75 mL
12-12 C44966AB
Explanation of restrictions (if fluidic self-check is not enabled):

Number of times used: 1-20
4 Select Detail. The Replace Sheath Fluid Filter window appears.
Select Replace. A pop-up window appears to reset the maintenance date as the current date that
5 Select Detail. The Replace Peristaltic Pump Tubing window appears.
C44966AB 12-13
Select Replace. A pop-up window appears to reset the maintenance date as the current date that
Adding the Deep Clean Solution
Check occasionally whether the Deep Clean solution in the Deep Clean solution bottle is sufficient.
Replace the Deep Clean solution when the maintenance reminder prompts you to do so.
WARNING
Risk of chemical injury from Contrad 70® reagent. To avoid contact with the
Contrad 70® reagent, use barrier protection, including protective eyewear,
gloves, and suitable laboratory attire. Refer to the Safety Data Sheet for details
about chemical exposure before using the chemical.
1 Make 60 mL of Deep Clean solution by mixing 30 mL of Contrad 70® and 30 mL DI water in the
Deep Clean solution bottle and swirl the solution gently to create the Deep Clean solution.
2 Verify that the Cytometer is in standby state or is turned off.
3 Remove the right-side cover of the instrument. Refer to Right-Side Cover Removal and
Reinstallation.
12-14 C44966AB
4 Remove the Deep Clean solution bottle and open the cap.
5 Add 60 mL Deep Clean solution to fill the bottle to the shoulder.
C44966AB 12-15
6 Tighten the cap, and attach the Deep Clean solution bottle to the bracket.
7 Install the right-side cover (see Right-Side Cover Removal and Reinstallation), and fasten the
thumbscrews.
8 Reset the maintenance reminder tracker. Refer to Managing the Maintenance Reminder.
Replacing the Sheath Fluid Filter
It is recommended to replace the sheath fluid filter every six months when the maintenance
reminder prompts you to do so. The life of the filter is related to the quality of the sheath fluid used.
If it is found that there are impurities in the light scatter pattern, replace the sheath fluid filter.
1 Select Standby on the left of the screen to place the instrument in standby state, or shut off the
Cytometer's power.
2 Remove the right-side cover. Refer to Right-Side Cover Removal and Reinstallation.
12-16 C44966AB
3 Press the spring piece on the quick connector and the Cytometer on the upper side of the filter,
and disconnect the quick connector.
4 Repeat Step 3 for the quick connector behind the quick connector removed in the previous
step.
NOTE The spring is on the opposite side.
5 Remove the sheath fluid filter from the bracket.
6 Connect the new, unused filter using the quick connector springs.
C44966AB 12-17
7 Attach the filter to the filter bracket.
8 Remove and set aside the vent cap.
9 Turn on the Cytometer and open the software.
10 Select Prime in the Cytometer menu.
12-18 C44966AB
WARNING
If the vent cap is not sealed tightly, unstable flow rate of the sheath fluid can
result, and leakage of the sheath fluid can occur.
11 Observe the liquid level in the filter during the prime cycle. When the liquid level reaches the
upper section of the filter, reinstall the vent cap to prevent air leakage.
12 Install the right-side cover (see Right-Side Cover Removal and Reinstallation), and lock the
screw.
13 If the problem persists, contact us.
14 Run the system startup program. Refer to Running the System Startup Program [Without
Autoloader] or Running the System Startup Program [With Autoloader] in CHAPTER 4, Daily
Startup.
15 Reset the maintenance reminder tracker. Refer to Managing the Maintenance Reminder.
C44966AB 12-19
Replacing the Sample Probe and/or the Sample Peristaltic Pump Tubing
WARNING
Risk of biohazardous contamination if you have skin contact with the sample
probe or the sample peristaltic pump tubing. The sample probe and the sample
peristaltic pump tubing could contain residual biological material and must be
handled with care. Clean up spills immediately. Dispose of the sample probe and
the sample peristaltic pump tubing in accordance with your local regulations and
It is recommended to replace the tubing of the sample peristaltic pump every six months, as tubing
used for an excessively long time can cause degradation of the stability of the sample flow and
increase of the CV detected.
[DxFLEX without Autoloader]
1 Place the instrument in standby.
2 Remove the right-side cover. Refer to Right-Side Cover Removal and Reinstallation.
3 Remove the front cover. Refer to Front Cover Removal.
4 Remove the sample pump cover thumbscrew and the sample pump cover.
12-20 C44966AB
5 Take out the sample peristaltic pump tubing.
6 Remove the flow cell PEEK tubing from the sample peristaltic pump tubing .
C44966AB 12-21
7 Lift the sample probe out of the wash station.
8 Remove the sample peristaltic pump tubing from the sample probe.
9 Dispose of the old sample probe and/or sample peristaltic pump tubing in accordance with
your local regulations and acceptable laboratory procedures.
12-22 C44966AB
10 Connect the sample peristaltic pump tubing to the sample probe.
11 Connect the PEEK tubing to the sample peristaltic pump tubing.
C44966AB 12-23
12 Slide the sample probe into the wash station.
13 Install the sample peristaltic pump tubing, taking care not to use any sharp tools, ensuring that
the tube is fully inserted into the groove.
NOTE Use the sample pump cover thumbscrew to insert the sample peristaltic pump tubing into the
groove while manually rotating the sample pump.
12-24 C44966AB
IMPORTANT To ensure the proper placement of the sample probe, push the sample pump cover up while
installing the thumbscrew.
15 Install the right-side cover (see Right-Side Cover Removal and Reinstallation), and lock with the
screw.
16 Install the front cover. Refer to Front Cover Removal.
[DxFLEX with Autoloader]
1 Place the instrument in standby.
2 Remove the top cover of the autoloader and open the front autoloader door. Refer to Top and
Front Cover of DxFLEX Autoloader Removal and Reinstallation.
3 Remove the front cover. Refer to Front Cover Removal.
C44966AB 12-25
4 Remove the sample pump cover thumbscrew and the sample pump cover.
5 Remove the sample PEEK tubings from the sample peristaltic pump tubing.
12-26 C44966AB
6 Take out the sample peristaltic pump tubing.
7 Loosen the sample probe set screw.
C44966AB 12-27
8 Pull the sample PEEK tubing down and out of the clip.
9 Pull the sample PEEK tubing through the routing holes.
12-28 C44966AB
10 Lift the sample probe out of the wash station.
11 Dispose of the old sample probe and/or sample peristaltic pump tubing in accordance with
your local regulations and acceptable laboratory procedures.
12 Insert the sample probe into the probe holder and wash station.
C44966AB 12-29
13 Route the sample probe PEEK tubing through the routing holes.
14 Clip the sample PEEK tubing in place.
12-30 C44966AB
15 Ensure the sample probe block is flush with the probe bracket (1) then tighten the sample probe
set screw (2).
c
b
16 Connect both the sample PEEK tubing and flow cell PEEK tubing to the sample peristaltic pump
tubing.
C44966AB 12-31
17 Install the sample peristaltic pump tubing, taking care not to use any sharp tools, ensuring that
the tube is fully inserted into the groove.
NOTE Use the sample pump cover thumbscrew to insert the sample peristaltic pump tubing into the
groove while manually rotating the sample pump.
IMPORTANT To ensure the proper placement of the sample probe, push the sample pump cover up while
installing the thumbscrew.
19 Install the top cover of the autoloader and close the front autoloader door. Refer to Top and
Front Cover of DxFLEX Autoloader Removal and Reinstallation.
12-32 C44966AB
20 Install the front cover. Refer to Front Cover Removal.
Inspecting the Liquid Flow Path for Leaks
WARNING
The liquid flow tubing can aged and cracked or the connector can be loosened.
Liquid leakage can lead to biological harm. To reduce occurrence of such
problems, carry out liquid flow tubing inspection every six months and ensure that
the Fluidics module functions without any leaks. If any leaks are found when using
the Cytometer, stop the experiment immediately and look for the source of the
leak.
1 Remove the right-side cover of the instrument. Refer to Right-Side Cover Removal and
Reinstallation.
2 Perform instrument initialization to enable the sheath fluid to flow. Refer to Initializing the
3 Check the connectors and tubes in the Fluidics module, and check whether any liquid leaks out.
4 Check the sheath fluid, backflush, and waste liquid connector on the back of the Cytometer, and
check whether any liquid leaks out.
5 Place the instrument in standby state, complete the priming procedure, and check whether the
Fluidics module has any liquid leakage.
6 If any liquid leaks out and the point of liquid leakage is from the filter, try to tighten the filter
connector and check again. If the problem persists, replace the sheath fluid filter.
7 If any liquid leaks out from any other connector or tube, stop running the instrument and
contact us.
C44966AB 12-33
Priming the Flow Cell

Priming of the flow cell is required when:
• The instrument is not used for a long period of time.

• The sheath fluid is refilled.
• The instrument is being used for the first time.
• The signal of the instrument is poor or the signal drifts.
• The sheath filter is replaced.
1 Ensure that the instrument is in standby state.
NOTE If the instrument is not already in the standby state, select Standby from the Cytometer Menu or
select Standby in the Data Acquisition Control screen.
2 Select Prime from the Cytometer Menu to prime the flow cell. Wait for the beep and for the
Instructions window to close.
Otherwise, look for the status bar to display that priming has completed.
3 Run Daily Clean to clean the sample line. Refer to Daily Clean in CHAPTER 11, Cleaning
Procedures.
12-34 C44966AB
Nonscheduled Replacement/Adjustment 12
Changing the Event Rate Setting

The Event Rate Setting adjusts the collection setting around signal measurement so that the system
is able to optimize the acquisition of events ensuring optimal sensitivity and abort rates when
acquiring at higher event rates.
1 Select Event Rate Setting in the Advanced menu. The Event Rate Setting window appears.
2 Select High if the event rate is >10,000 events/second.

Or
Select Default if the event rate is <10,000 events/second.
3 Select OK.
Nonscheduled Replacement/Adjustment
Calibrating the Sample Flow Rate
Calibrate the sample flow rate:
• After replacing the sample peristaltic pump tubing.

• If a precise volumetric measurement is required.
The accuracy of concentration calculations can be affected by the sample flow rate.
Refer to Calibrating the Sample Flow Rate in Plate Mode in APPENDIX C, DxFLEX [With Autoloader
-Plate Mode] for instructions on calibrating the Sample Flow Rate in the Plate Mode.
1 Verify that the instrument is in the initialized state.
C44966AB 12-35
2 Select Calibrate Sample Flow Rate in the Cytometer menu.
3 Select the flow rate to be calibrated.

If you want to calibrate all flow rates at once, fast calibration is recommended. The rate selected
in the Calibrate Sample Flow Rate window overrides the rate selected in the Acquisition
window.
12-36 C44966AB
4 Prepare one sample tube with 1 mL of clean deionized water then use a calibrated analytical
balance to measure the weight of the prepared sample tube. Record the weight and enter it into
the software.
Without Autoloader
With Autoloader (Single Tube Mode or Carousel Mode)
NOTE DxFLEX [With Autoloader]: Select in the Calibrate Sample Flow Rate window
to set the tube location and the Carrier Type.
Input weight: 0-100
Collection time (score): 1-30
5 Select Next and put the sample tube in the sample loading position (see Figure 1.11, Figure 1.12).
C44966AB 12-37
6 Select Initialize to start the sample run.
Without Autoloader
7 Select Run to begin acquiring the sample.
Without Autoloader
12-38 C44966AB
8 Wait for the sample run to finish, remove the sample tube, and use the analytical balance to
measure the weight and record the value.
9 Select Next to determine if the results fall within the acceptable range.
If the results fall within the acceptable range, the current setting is kept.
C44966AB 12-39
If deviation occurs, the setting is automatically corrected.
NOTE Select [According to this value calibrate other flow rate] to apply the coefficient directly to
skip calibrating the Medium and Slow flow rate.
Setting Laser Delay

Laser delay is preset for QC. Only change the laser delay if the software prompts you that there is a
difference in the actual delay and the default delay.
1 Select Delay Setting from the Advanced menu. The Delay Setting window appears.
Current delay (μs): -160-160
12-40 C44966AB
2 Set the current to the actual delay for the specified detector stated in the error message
received.
3 Select Set as Default.
4 Select Close.
Replacing the Optical Filter

When the optical filter is damaged or it is required to use an optical filter with a non-standard
wavelength, you must replace the optical filter yourself. For the specific part number of the optical
filter, consult your Beckman Coulter Representative or your local dealer.
1 Confirm that the instrument is in the standby state or that the instrument is turned off.
2 Confirm the laser corresponds to the channel in which the optical filter is to be replaced.
3 Open the top cover of the instrument.
4 Press the spring piece of the WDM cap corresponding to the laser, and open the WDM cap.
C44966AB 12-41
CAUTION
Risk of damage to the optical filter. Do not touch the optical filter glass piece.
Touching the optical filter glass piece can obscure and/or scratch the optical filter
glass piece.
5 Use vertical force to remove the optical filter to be replaced, and note the color identification
and wavelength identification on the optical filter bracket.
6 Insert the optical filter to be installed vertically into the corresponding position, taking care to
align the wavelength identification with the left, and that the bracket is inserted into the
bottom.
7 Close the WDM cap and the instrument top cover.
8 Turn on the Cytometer and open the software.
12-42 C44966AB
9 Select Detector Configuration in the Cytometer menu and create a new instrument
configuration based on the settings of the new optical filter. Refer to Verifying, Selecting,
Editing, and Creating Detector Configuration in CHAPTER 6, Data Acquisition and Sample
Analysis.
Set this new configuration as the current configuration.
Replacing the Fuse

Use a 5 A, time delay, T 5 AL, 250 VAC fuse.
WARNING
Risk of personal injury. A shock hazard exists if the power cable is connected. Turn
off the Cytometer and disconnect the primary power cable before performing
these procedures.
1 Turn off the Cytometer, and disconnect the power cable.
C44966AB 12-43
2 Press both sides of the fuse holder of the instrument inwards using a flat head screwdriver, and
pull out the fuse holder.
IMPORTANT Select well-performing products that comply with the specifications required, to ensure that
the instrument can function normally and safely.
3 Check whether the fuse installed is blown, and replace the blown fuse with a new one.
The specifications of the fuse required are: T 5 AL 250 VAC, delay blow fuse, 5A, 250 VAC,
5 x 20 mm. Beckman Coulter recommends using SCHURTER 0034.3124.
4 Insert the fuse holder.
12-44 C44966AB
5 Reconnect the power cable.
Replacing the Sheath Fluid Harness and/or Waste Harness
Replace the sheath fluid harness and/or the waste harness if you have a faulty sheath fluid sensor
and/or waste sensor.
2 Remove sheath and/or waste pickup tubing from the appropriate container.
3 Disconnect the blue harness (1) from the sheath fluid container (2) and/or the yellow harness
(3) from the waste container (4) from the fluid connector panel (5) on the back right corner of
the instrument according to the color code.
d
b
MANUFACTURED
upper upper
REF
SN
EC REP
IVD c e
lower
C44966AB 12-45
Disconnect the waste (1) and/or sheath (2) level sensors.
b c
4 Dispose of the sheath fluid harness and/or the waste harness according to your laboratory
procedures.
5 Insert the new sheath and/or waste pickup tubing into the appropriate container.
6 Connect the blue harness (1) from the sheath fluid container (2) and/or the yellow harness (3)
from the waste container (4) to the fluid connector panel (5) on the back right corner of the
instrument according to the color code.
d
b
MANUFACTURED
upper upper
REF
SN
EC REP
IVD c e
lower
12-46 C44966AB
Connect the waste (1) and/or sheath (2) level sensors.
b c
C44966AB 12-47
Changing Sample Mixing and Backflush Settings
WARNING
Risk of biohazardous contamination. Enabling sample mixing for 1.5-mL and 2-mL
sample tubes in the semi-automatic sample injection mode can result in sample
splashing. Exceeding 300-μL sample volume when using 1.5-mL and/or 2-mL
sample tubes can also result in sample splashing. Disable sample mixing in the
semi-automatic sample injection mode when using 1.5-mL and 2-mL sample tubes
and do not exceed 300-μL sample volume.
WARNING
Risk of biohazardous contamination. Exceeding 100-μL sample volume when using
plates with the mix setting set to high intensity could result in sample splashing.
Use a lower mix intensity for samples volumes over 100-μL and do not exceed
200-μL sample volume.
WARNING
Risk of biohazardous contamination. Exceeding 2-mL sample volume when using
12x75 mm tubes in the DxFLEX Autoloader carousel could result in sample
splashing. Do not exceed 2-mL sample volume in 12x75 mm tubes on the DxFLEX
Autoloader carousel.
The sample mixer can be enabled or disabled if necessary. The sample mixing duration can also be
increased or decreased if necessary.
Whenever a sample is likely to leave residue or cause contamination, the backflush time can be
increased to reduce cross contamination.
IMPORTANT Mix and Backflush settings can be changed on the DxFLEX [With Autoloader] in the Carrier tab
of the Options window.
1 Open the CytExpert for DxFLEX software and confirm that the instrument is connected. Refer
to Opening the Software in CHAPTER 4, Daily Startup.
12-48 C44966AB
2 Select Cytometer Configuration in the Cytometer menu. The Cytometer Configuration window
appears.
[DxFLEX Without Autoloader]
[DxFLEX With Autoloader]
C44966AB 12-49
IMPORTANT Sample mixing cannot be changed from the Cytometer Configuration window on the DxFLEX
[With Autoloader]. To change mix setting, refer to Creating Experiments in CHAPTER 6, Data Acquisition
and Sample Analysis.
3 [DxFLEX Without Autoloader Only]: Select the Sample Mixing check box to enable sample
mixing.
Or
Deselect the Sample Mixing check box to disable sample mixing.
4 [DxFLEX Without Autoloader Only]: Change the sample mixing duration to the desired time.
NOTE The default setting is 1 second. Select Default to set the Cytometer configuration settings back
to the factory default settings.
5 Change the backflush duration to the desired time.
NOTE The default setting is 3 seconds. Select Default to set the cytometer configuration settings back
to the factory default settings.
6 Select OK.
Calibrating the Carrier [DxFLEX With Autoloader]

Use the following procedure to calibrate the carrier position:
• Upon installation
• After a new carrier type, carousel type, or plate type is defined for first use
• After changing plate manufacturers of the same previously calibrated plate
• When optimizing carrier performance
Refer to Calibrating the Carrier in Plate Mode in APPENDIX C, DxFLEX [With Autoloader -Plate
Mode] for instructions on calibrating the carrier using the Plate Mode.
Calibrating the Carrier in Single Tube or Carousel Mode
1 Ensure the sample injection control mode is set to Single Tube or Carousel Mode. Refer to
Selecting the Proper Sample Injection Mode [with Autoloader] in CHAPTER 4, Daily Startup.
2 If the instrument is in initialized state, select Standby.
12-50 C44966AB
3 Select Advanced > Calibrate Carrier. The Calibrate Carrier window appears.
4 Select carousel and place the carousel inside the autoloader. Refer to Installing a Carousel in
CHAPTER 1, System Overview.
5 Place an empty tube in the carousel tube location 1 and leave the autoloader cover open.
6 Select Calibrate. The message Please confirm that the correct carrier is placed properly and press OK.
appears. Select OK.
C44966AB 12-51
CAUTION
Risk of instrument damage. Do not crash the probe into the bottom of the tube as
this will cause irreparable damage to the probe. Move the probe one step at a time
when lowering the probe on the Z-axis toward the bottom of the carousel. You will
hear a click when the probe makes contact with the bottom of the tube. Do not
move the probe any lower after you hear the click. Move the probe back up three
steps after you hear the click, this is the correct position for the Z-axis of the
probe.
7 Select and to adjust the sample probe positions in the Z-axes.

Ensure the sample probe touches the bottom of the tube.
NOTE The sample probe should only make contact with the bottom of the tube.
NOTE The Z-axis arrows moves the sample probe up and down.
8 Select Test to verify the sample probe position.

Listen for the click to ensure the probe has made contact with the bottom of the tube. Do not
move the probe any lower after you hear the click. Move the probe back up three steps after
you hear the click; this is the correct position for the Z-axis of the probe.
9 Select Next.
10 Select Finish to save the settings.
11 Select Close to quit the program.
12-52 C44966AB
APPENDIX A
Instrument Installation
Overview
The DxFLEX is installed by your Beckman Coulter Service Representative. Do not open the box or
crate. Wait for a qualified Beckman Coulter Service Representative.
Validate the Install the

Ô Open the package Ô Ô Prepare to boot
environment instrument
• Instrument Transportation and Storage

• Installation Environment Validation
• Unpacking the Instrument and Inspecting the Materials for Defects or Omissions
• Updating and Reinstalling the Software
Instrument Transportation and Storage
Refer to Preparing the Instrument for Transport or Storage in CHAPTER 11, Cleaning Procedures,
prior to transportation or storage.
Attention to the following items is required when transporting or storing the instrument:
• Take caution to protect the instrument from exposure to rain or sunlight.
• Always place the instrument on a flat, stable surface, and take note of the symbol for this side up.
• Temperature range: -10°C~55°C; Humidity: 80%
• To prevent extrusion, the load on top cannot exceed 100 kg.
• Cytometer net weight 23 kg, gross weight 27 kg; transport the instrument using only
appropriate equipment so as to guard against personal injury.
Installation Environment Validation
IMPORTANT This instrument is intended for indoor use only.

Verify whether the installation environment satisfies the following requirements:
C44966AB A-1
Installation Environment Validation
Worktable
CAUTION
Risk of instrument damage. Place the instrument on a level surface. Failing to do
so places the system is in danger of toppling and can result in damage. Take all
necessary precautions throughout the process of storing or transporting the
instrument.
• The tabletop must be smooth and level.

• Minimum tabletop load bearing capacity: 50 kg.
• The tabletop must not vibrate or shake.
• Minimum tabletop dimensions: 120 cm x 80 cm; minimum vertical space above tabletop: 80 cm
• Position the instrument in such a manner that it will facilitate disconnection of the power cable
at the instrument end.
Ventilation and Cleaning

IMPORTANT If necessary, use ventilation equipment, but airflow must not be allowed to blow directly on
the system, as it can affect the reliability of the data.
• Ensure that the working environment is well ventilated for proper heat dissipation.
• Maintain a clearance of at least 20 cm from the back of the instrument for heat dissipation.
• Keep the environment as dust free as possible.
• Avoid direct exposure to sunlight.
• Avoid placing near heat sources or exposing to drafts.
• Avoid corrosives or flammable gases.
A-2 C44966AB
Installation Environment Validation A
Power Source
DANGER
Risk of electric shock and/or instrument damage. Ensure that the power source is
properly grounded. Improper grounding can cause electric shock and damage the
system. Verify that the output voltage of the power outlet conforms to the system
requirements and that a 5 A, time delay, T 5 AL 250 VAC fuse is installed. To
prevent personal injury, Beckman Coulter recommends using a power source
designed to protect against electrical shock.
CAUTION
Possible instrument damage could occur if you use an extension cord or a power
strip to connect the Cytometer. Always plug the Cytometer into a dedicated outlet
with an isolated ground.
The power source requirements are as follows:
• This instrument has been tested to and meets all applicable requirements for CE Marking.
• This instrument complies with the emission and immunity requirements described in IEC
61326-1 and 61326-2-6 series.
• This equipment has been designated and tested to CISPR 11 Class A. In a domestic environment
it may cause radio interference, in which case, you may need to take measures to mitigate the
interference.
• It is advised that the electromagnetic environment should be evaluated prior to operation of
the device.
• Do not use this device in close proximity to sources of strong electromagnetic radiation
(unshielded intentional RF sources), as these may interfere with the proper operation.
• 100-240 volts, 50/60 Hz, 3-wire power cable, well grounded.
• Amperage not less than 10 A.
• The system requires a well-grounded power outlet (150 VA normal, 250 VA max) to provide the
necessary power.
• Distance from system to socket less than 1.5 m.
Temperature and Humidity
CAUTION
Risk of instrument damage and/or erroneous results. To ensure reliability, the
system must be operated in the specified environment, within the required
temperature and humidity ranges. If the ambient temperature or humidity level
falls outside the ranges mentioned above, use appropriate air conditioning.
• Ambient temperature: 15-30°C with fluctuations of no more than <±2°C per hour.
• Relative humidity: 15%-80%, non-condensing.
C44966AB A-3
Unpacking the Instrument and Inspecting the Materials for Defects or Omissions
Waste Disposal
WARNING
associated tubing might contain residual biological material and must be handled
procedures.
Dispose of the system’s waste in accordance with your local regulations and acceptable laboratory
procedures.
Unpacking the Instrument and Inspecting the Materials for Defects or

Omissions
Take care to store the instrument in a suitable environment where it can remain in the proper
position.
Check that the following components on the packing list are present:
• Cytometer
• Cables
• Sheath fluid container
• Waste container
• Sheath fluid tubing
• Waste tubing
• USB configuration key
• Software USB
If you have a DxFLEX [With Autoloader], you will also receive the following components:
• Two carousels
• Plate adapter (optional)
• Right cover assembly
• Bracket assembly
• Bottom plate enhancement
• Accessory box
A-4 C44966AB
Updating and Reinstalling the Software A
Installing the Instrument and Connecting the Equipment [DxFLEX Without

Autoloader]
The DxFLEX must be installed by a trained Beckman Coulter service engineer.
Use the appropriate power cable plug for your geographic region.
For connecting the equipment, refer to CHAPTER 1, System Overview.
Updating and Reinstalling the Software
The installation process workflow is as follows:
Update or reinstall the

Uninstall the CytExpert for
Ô CytExpert for DxFLEX Ô Start the software
DxFLEX software
software
The CytExpert for DxFLEX software can be installed on a computer that meets the minimum
specifications (see Instrument Specifications in CHAPTER 1, System Overview) for analysis-only
use.
Required materials
The following materials are required to install the CytExpert for DxFLEX software:
• DxFLEX flow cytometer.
• DxFLEX Workstation.
• CytExpert for DxFLEX software installation USB.
• Authorized Beckman Coulter USB configuration key.
Uninstalling the Software

IMPORTANT Ensure you backup all of your data before uninstalling the software.
1 Go to the “Control Panel” and find the “Program Uninstall” page.
C44966AB A-5
IMPORTANT Do not remove the existing settings, database, Cytometer configuration and temporary files.
2 Select the “CytExpert for DxFLEX ”program, right-click to find “Uninstall” and follow the
instructions to uninstall.
Updating and Reinstalling the CytExpert for DxFLEX Software
1 Insert the software USB into the computer.
NOTE If the AutoPlay window appears, select Open folder to view files.
A-6 C44966AB
2 Select CytExpert for DxFLEX 2.0 Setup.exe. The CytExpert Setup Welcome window appears.
A User Account Control message may display the message Do you want to allow the following
program from an unknown publisher to make changes to this computer. Select Yes to continue.
3 Select Next.
4 Select all three support program check boxes in the CytExpert Setup Prerequisites window.
C44966AB A-7
5 Select Next. The Welcome to CytExpert Setup Wizard window appears.
6 Select Next. The Beckman Coulter License Terms window appears.
7 Read the Beckman Coulter Customer End User License Agreement.
A-8 C44966AB
8 Select the I accept the terms of this agreement check box.
NOTE The checkbox is not selectable until you scroll all the way to the end of the agreement.
9 Select Next. The Choose a file location window appears.
10 Select Next. The Begin installation of CytExpert window appears.
C44966AB A-9
11 Select Install to begin installing the software. The Installation Progress window appears.
NOTE The software will install into the default file path provided unless otherwise specified.
12 A Windows Security window may appear. Select Install this driver software anyway to install the
USB drive.
13 The following software prompt appears.
Select OK.
A-10 C44966AB
14 Wait for the software to finish installing. The install complete window appears.
15 Select Close to finish the CytExpert for DxFLEX software installation.
Starting the Software
1 Insert the USB configuration key into the USB port of the computer.
2 Start the software. Refer to Opening the Software in CHAPTER 4, Daily Startup, for detailed
instructions on opening the software and confirming the connection status.
NOTE If the software shows Connected, data collection and analysis can be completed.
C44966AB A-11
A-12 C44966AB
APPENDIX B
Barcode Specifications for the DxFLEX
[With Autoloader]
Barcode Sample Identification
Barcode symbols are a highly accurate and efficient procedure for identifying and processing
laboratory samples. Beckman Coulter instruments use four barcode symbologies (types) to identify
specimens:
• Code 128
• Code 39
• Codabar
• Interleaved 2-of-5.
The barcode reader senses the difference between enabled barcode symbologies in a run.
CAUTION
A misread label can cause one sample ID to be read as another sample ID. The
laboratory’s process for printing, placing, and meeting all barcode specifications
is important to achieve highly accurate reading. Follow the barcode specifications
to avoid inaccurate reading of the barcode label.
Figure B.1 Barcode Label

b
c
ABC123456789 d
1 Quiet zone
2 Barcode symbol
3 Sample ID
Correct Placement of The Barcode Label
The barcode label must be placed a minimum of 25.4 mm (1.0 in.) from the top of the tube. Refer to
Figure B.2.
C44966AB B-1
Barcode Specifications for the DxFLEX [With Autoloader]
Barcode Label Specifications
Figure B.2 Barcode Label Placement
>20mm
<7.5°
1 20 mm (0.79 in.) minimum

2 <7.5 degrees
Put labels on the tubes so that the bars follow one another in a vertical sequence. Refer to Figure B.2.
The barcode reader scans the tube vertically. Do not tilt the label more than 7.5 degrees from the
axis of the tube.
Put the tubes in the carousel so that the barcode symbols are visible through the slots in the front
of the carousel. When viewed at eye level, the full symbol, including the quiet zones, must be visible
through the slot and above the bottom of the carousel.
NOTE The DxFLEX [With Autoloader] Flow Cytometry System rotates the tube as needed so the barcode
label can be read.
Barcode Label Specifications
CAUTION
A misread label can cause one sample ID to be read as another sample ID. The
laboratory’s process for printing, placing, and meeting all barcode specifications
is important to achieve highly accurate reading. Follow the barcode label
specifications to keep the rate of misread labels to a minimum.
The quality of the barcode symbol and the label is important for accurate reading. For high
accuracy, use labels that meet all of the specifications.
When possible, print the sample ID on the label in alphanumeric characters so the operator can
manually enter the barcode information if the barcode symbol cannot be read.
B-2 C44966AB
Barcode Label Specifications B
Label Size and Thickness

The length of the label must be less than 38 mm (1.50 in.). The label includes the barcode symbol
and a minimum quiet zone of 3.5 mm (0.14 in.) at each end of the symbol. Refer to Figure B.3.
Figure B.3 Barcode Label Specifications

b b
ABC123456789 c
d
1 Quiet zones 3.5 mm (0.14 in.) minimum
2 Barcode symbol height 7 mm (0.28 in.) minimum
3 Barcode label length 38 mm (1.50 in.) maximum
The width of the barcode label must be less than half the circumference of the sample tube.
Label thickness, including adhesive, must be 0.09 mm (0.0036 in.) maximum. Total thickness for all
labels and adhesives put together must be 0.36 mm (0.0144 in.) maximum.
Symbol Dimensions
The height of the barcode symbol must be 7 mm (0.28 in.) minimum.
See Table B.2.
Label and Print Quality

All barcode symbols must agree with the American Identification Manufacturer’s (AIM) Uniform
Symbology Specification.1
All barcode symbols must be printed at print quality class “B” or better as defined by the American
National Standards Institute (ANSI).2 Several factors affect print quality:
• Labels must be clean, not yellowed, and used before the expiration date.
• Print the barcode symbol on material that is reflective and has a matte finish. Use a background
diffuse reflectance of 80% or more for maximum contrast.
• The labels must not have defects such as spots, lines, missing sections, cuts, folds, or density
problems.
• The bars in the barcode symbol must be well-defined. Edges must be constant (not irregular),
so the bars and spaces have the correct widths for the barcode symbology used.
C44966AB B-3
Barcode Error Rate
Barcode Error Rate
CAUTION
A misread label can cause one sample ID to be read as another sample ID.
Whenever possible use a barcode symbology and configuration choices that
provide the most accurate barcode reading.
The quality of the barcode symbol and the label is important for accurate reading. To get the highest
possible accuracy, only use labels that meet all the specifications described for labels and symbols.
Deviations from these specifications make the barcode more difficult to read and allow for a
possible increase in the error rate.
The symbology and the configurable parameters that the laboratory selects have an effect on the
error rate. Certain features of the symbologies and the selections made by the laboratory have an
important effect on the accuracy of the barcode reading system. In general:
• Code 128 and Code 39 are more accurate and have lower error rates than Codabar or Interleaved
2-of-5.
• CLSI recommends Code 128 because of its accuracy, compact form, and self-checking
capabilities.3
• A checksum greatly increases accuracy. Use a checksum with Interleaved 2-of-5 and Codabar
because they are less accurate symbologies.
• Select the fixed length option, if available, because it is more accurate than the variable length
option.
• To keep label and printing flaws to a minimum, use a narrow element of more than 0.15 mm
(0.006 in.).
Beckman Coulter recommends the use of:
• Code 128
• Checksum for all other symbologies
• Fixed length code symbols
• Narrow bar sizes of 0.15 mm (0.006 in.) minimum.
Barcode Symbologies
Beckman Coulter instruments use four barcode symbologies for specimen identification, see
Table B.1.Within the given specifications, the MCL reader and the optional handheld barcode
reader automatically distinguish the following barcodes:
B-4 C44966AB
Barcode Labels B
Table B.1 Barcode Symbologies
Barcode Type Description

Code 128 • Variable length
(also known as USD-6) • ASCII 32 to ASCII126
• Self-checking
• Continuous code; intercharacter space is part of code structure for higher
density of code per square inch; compact barcode
• Code 128 is recommended by the Clinical and Laboratory Standards Institute
(CLSI) for its accuracy, compact form, and self-checking capabilities3
• Code 128B - Maximum 8 alphanumeric characters / Minimum 3 numeric
characters
• Code 128C - Maximum 16 numeric characters / Minimum 3 numeric
characters
(The use of 15 numeric characters is invalid)
• The following character is not allowed: !
Code 39 • Variable length
(also known as 3-of-9 • Includes 43 data characters; 26 letters (uppercase A-Z), 10 digits (0-9), six
and USD-3) symbols (. $ / + % -) and a space
• Strong self-checking properties
• Checksum
• Discrete code; white spaces are not part of this code
• Maximum 7 characters
(6 data characters + 1 checksum character).
Interleaved 2-of-5 • Numerics only
(also known as I2 of 5, • Checksum
USD-1, and USD-1.25) • Lower density of code per square inch; longer label
• Requires an even number of digits to be encoded, a leading “0” must be
added if the number count is odd
• Fixed 14 characters
Codabar • Variable length
(also known as USD-4 • Includes 16 data characters; 10 digits (0-9), and six symbols (. $ / + % -)
and NW7) • Has specific start and stop characters which lead to improvement in
readability
• Checksum
• Lower density of code per square inch; longer barcode
• Maximum 10 characters
Barcode Labels
A barcode consists of black lines (bars) and white lines (spaces), which are called elements.
C44966AB B-5
Autoloader Barcode Reader
There are narrow elements (NE) and wide elements (WE). The barcode symbology determines their
arrangement.
CAUTION
Sample misidentification can occur from the use of incorrect, poor quality,
damaged, dirty or improperly placed barcode labels. Follow the specifications in
this section to create your barcode labels to prevent incorrect sample
identification. See also Putting a Barcode Label on a Sample Tube.
The instrument supports preprinted labels.
Barcode Label Optical Characteristics at 650 nm ±10%

• Print Contrast Signal (PCS): 80% minimum.
• Reflectivity of Media (RW): 80% minimum.
• Reflectivity of Ink (RB): 16% maximum.
• No spots or voids; no ink smearing.
• Edge roughness is included in the bar and space tolerances.
RW – RB
PCS = -------------------  100%
RW
Table B.2 Code-Related Specifications
Code Interleaved Codabar* Code 39* Code 128*

2-of-5*
Narrow element (NE) width 0.15 mm 0.15 mm 0.15 mm 0.15 mm
Wide element/narrow 3:1 N/A 3:1 N/A
element ratio (WE/NE)
Intercharacter gap No 0.010 in. minimum _NE No
Data digits 14** 1 to 10** 1 to 7** 2 to 16
* See AIM Uniform Symbology Specification, Rev. 1995 for detailed specification.
** Includes checksum character
Autoloader Barcode Reader
The Autoloader barcode reader uses a visible-laser type reader containing a Class II laser, operating
at 650 ±10 nm, with a maximum power output of 1 mW.
B-6 C44966AB
Checksum Algorithm B
Checksum Algorithm
CAUTION
Use of barcodes is an extremely accurate and effective method of positive patient
identification. Certain features, such as checksum digits, maximize accuracy in
reading Codabar, Code 39 and Interleaved 2-of-5 labels. In one study, the use of
checksum digits detected 97% of misread errors.
Use checksums to provide protection against occasional misread errors caused by
problems such as damaged or misapplied labels. If you must use barcodes without
checksums, Beckman Coulter recommends that you verify each barcode reading
to assure correct patient identification.
Beckman Coulter strongly recommends the use of barcode checksums to provide automatic checks
for read accuracy.
C44966AB B-7
Checksum Algorithm
B-8 C44966AB
APPENDIX C
DxFLEX [With Autoloader -Plate Mode]
Overview
IMPORTANT The plate adapter is not validated for IVD use with the DuraClone B27 Reagent.
This chapter provides procedures for using the plate adapter and the plate mode with the DxFLEX
[with Autoloader]. The plate adapter is an optional accessory that can be purchased separately. For
further descriptions regarding the DxFLEX [With Autoloader], refer to CHAPTER 1, System
Overview.
• Installing the Plate Adapter

• Removing the Plate Adapter
• Using the CytExpert for DxFLEX Software
• Selecting the Plate Sample Injection Mode [With Autoloader - Plate Mode]
• Instrument Quality Control and Standardization
• Data Acquisition and Sample Analysis
• Using Heat Maps
• Daily Clean [With Autoloader - Plate Mode]
• Calibrating the Sample Flow Rate in Plate Mode
• Calibrating the Carrier in Plate Mode
The plate adapter supports three types of 96-well plates including:
• U-bottom 96-well plate

• V-bottom 96-well plate
• Flat bottom 96-well plate
C44966AB C-1
IMPORTANT Ensure the adapter locks firmly into place.
2 Push the locking buttons in and align the plate adapter so the arrow aligns with the tab on the
carousel hub.
C-2 C44966AB
Installing the Plate Adapter C
3 Place the plate on the plate adapter.
NOTE Ensure plate well A1 aligns with position A1 on the plate adapter.
C44966AB C-3
IMPORTANT Ensure the adapter locks firmly into place.
2 Push the locking buttons in and pull the plate adapter straight up.
C-4 C44966AB
Removing the Plate Adapter C
C44966AB C-5
Collection
Standby state [Plate Mode] Initialized state [Plate Mode]
1. Acquisition control. Controls sample loading/unloading and data acquisition and recording.
2. Acquisition status. Displays such information as the acquisition rate (Events/Sec), cell count, duration,
and abort (%).
3. Acquisition conditions. Sets the necessary conditions for recording data.
4. Sample flow rate. Sets the acquisition rate for data collection.
NOTE High acquisition rate increases the abort rate and measurement CVs.
Refer to Collection in CHAPTER 2, Using the CytExpert for DxFLEX Software for the Collection states
in Single Tube Mode and Carousel Mode.
C-6 C44966AB
Using the CytExpert for DxFLEX Software C
Test Tubes
[Plate Mode]
C44966AB C-7
1. Test tube management controls. Manages sample tubes. Used to add, copy, or delete attributes,
open the tube property, and open the compensation matrix.
2. Test tube status indication. Displays a colored symbol in front of each tube indicating the status of
the tube processing.
• indicates that the tube data was not collected.
• indicates that the tube data was acquired by selecting Run but not saved, and the tube data
can be overwritten.
• indicates that tube data was reacquired by selecting Run.

• indicates that the tube data was recorded and this data cannot be overwritten.
• indicates that tube data was reacquired and recorded.

• indicates imported FCS data.
NOTE to the left of the test tube status indication symbol indicates that the sample has been
compensated.
• indicates the data file is missing or there is an error in the data file.
• indicates the Sample ID information does not match with the recorded tube barcode.
• indicates the data has been reviewed.
• indicates that tube data was not collected but was linked to Acq. Settings, and the
compensation value changes with the gain setting.
• indicates that tube data was not collected but was linked to Acq. Settings and was reviewed.
The compensation value changes with the gain setting
• indicates that tube data was not collected but was linked to Acq. Settings, and the
compensation value does not change with the gain setting.
• indicates that tube data was not collected but was linked to Acq. Settings and was reviewed.
The compensation value does not change with the gain setting.
• indicates that tube data was acquired by selecting Run and was analyzed .
• indicates that tube data was acquired by selecting Run and was reviewed.
• indicates that tube data was acquired by selecting Record or Auto Record, and was
analyzed.
• indicates that tube data was acquired by selecting Record or Auto Record, and was reviewed.
3. Test tube list. Displays the sample tubes used in the experiment. Right-click a tube in the list to
perform additional operations.
C-8 C44966AB
Using the CytExpert for DxFLEX Software C
New Experiment Test Tube Management Module for DxFLEX [With Autoloader - Plate Mode]
New Panel Experiment Test Tube Management Module for DxFLEX [With Autoloader - Plate Mode]
Refer to Test Tubes in CHAPTER 2, Using the CytExpert for DxFLEX Software for the Single Tube
Mode and Carousel Mode Test Tubes section description.
Software Settings
Refer to Software Settings in CHAPTER 2, Using the CytExpert for DxFLEX Software.
In the Carrier settings, you can set the following plate settings:
Plate Settings
• Plate type
• Sampling sequence
• Mix and backflush settings
C44966AB C-9
Selecting the Plate Sample Injection Mode [With Autoloader - Plate Mode]
Selecting the Plate Sample Injection Mode [With Autoloader - Plate

Mode]
Select Sample Injection Mode > Plate Mode in the Cytometer menu to change between the Single
Tube Mode, Carousel Mode, and Plate Mode. Refer to Selecting the Proper Sample Injection Mode
[with Autoloader] in CHAPTER 4, Daily Startup for instructions on selecting the Plate Mode.
The Plate Mode can be used for running small volumes using the following plates: 96-well
flat-bottom, 96-well V-bottom, and 96-well U-bottom.
Using Plate Mode
1 Select Sample Injection Mode > Plate Mode in the Cytometer menu to change the Sample
Injection mode selection.
2 The restart warning prompt appears on screen. Select OK.
3 Turn the Cytometer's main power switch off.
C-10 C44966AB
Selecting the Plate Sample Injection Mode [With Autoloader - Plate Mode] C
4 Turn the Cytometer’s main power switch on.

The sampler status icon located in the bottom right side of the screen changes to display Plate.
C44966AB C-11
7 Install the plate adapter.
8 Place the plate flat on the plate adapter and ensure that it is secure.
NOTE Ensure that plate well A1 aligns with position A1 on the plate holder.
9 If necessary, calibrate the carrier. Refer to Calibrating the Carrier [DxFLEX With Autoloader] in
C-12 C44966AB
Selecting the Proper Sample Injection Mode [with Autoloader]

Refer to Selecting the Proper Sample Injection Mode [with Autoloader] in CHAPTER 4, Daily Startup
for instructions on selecting the Plate Mode.
Running the System Startup Program [in Plate Mode]

The system startup program takes approximately 10 minutes.
NOTE The fluidic self-check feature is always enabled by default on all DxFLEX [With Autoloader]
instruments.
For instructions on running the System Startup Program in Single Tube Mode or Carousel Mode,
refer to Running the System Startup Program [With Autoloader] in CHAPTER 4, Daily Startup .
C44966AB C-13
The System Startup Program window appears.
System Startup Program Window in Plate Mode
C-14 C44966AB
The plate automatically sets the default well locations for the deionized water wells as shown
in the figure above. Between two and six deionized water wells must be specified to run the
System Startup Program.
NOTE To deselect wells, select the desired well and select Set As Empty.
NOTE To select new deionized water well locations, select the desired wells and select Deionized
Water.
The instrument begins to run the fluidic self-check. This process takes about 4 minutes. If you
are running in Plate Mode, the system message Please confirm that the correct plate is placed
properly and press OK appears. Select OK.
System Startup Program Step 2 in Plate Mode
After running the fluidic self-check, the system initializes again. The sample is loaded
automatically. This process takes about 3 minutes.
C44966AB C-15
The sample is unloaded after sample acquisition has finished. The system uses the remaining
time to warm up.
C-16 C44966AB
initialized.
C44966AB C-17
Refer to CHAPTER 5, Instrument Quality Control and Standardization for instructions.
Preparing the QC Well Plate
Required Materials
The following materials are required to complete the QC process:
• DxFLEX Daily QC Fluorospheres

• 96-well plate
— 96-well flat-bottom
— 96-well V-bottom
— 96-well U-bottom
• Vortexer
C-18 C44966AB
Instrument Quality Control and Standardization C
Preparation process
1 Take one 96-well plate and label one well as the QC sample well.
2 Use the vortexer or shake vigorously to thoroughly mix the bottle of DxFLEX Daily QC
Fluorospheres.
IMPORTANT Do not overfill the sample well.
3 Add 3-5 drops of DxFLEX Daily QC Fluorospheres to the sample well.
4 Place the well plate in the plate adapter.
Setting up the Plate
1 Select . The Carrier Settings window appears.
IMPORTANT Ensure the well position on the plate matches the well position selected in the software.
2 Select the appropriate QC well.
C44966AB C-19
3 Select the desired plate type from the Carrier Type dropdown menu.
4 Select the appropriate Mix and Backflush settings in the top of the Carrier Settings window.
NOTE If the mix intensity is set to high, the following warning appears:
C-20 C44966AB
Data Acquisition and Sample Analysis C
5 Select OK.
Collecting QC Data
For instructions on collecting QC Data, refer to Collecting QC Data in CHAPTER 5, Instrument Quality
Control and Standardization.
Refer to CHAPTER 6, Data Acquisition and Sample Analysis.
Setting Tube Property [With Autoloader-Plate Mode]

For instructions on setting the tube property in Single Tube Mode or Carousel Mode, refer to Setting
Tube Property [Single Tube or Carousel Mode] in CHAPTER 6, Data Acquisition and Sample Analysis.
1 [Standard Experiment]: Select to add tubes to the experiment. Skip to Step 3.
NOTE Use the dropdown to create multiple tubes.
Standard Experiment - Plate Mode Shown
Or
[Panel Experiment]: Select to add samples to the experiment. Proceed to Step 2.
NOTE Use the dropdown to create multiple tubes or to duplicate existing tubes without data.
Panel Experiment - Plate Mode Shown
C44966AB C-21
2 [Panel Experiment]: Right-click each sample and select New Tube to add tubes to each sample.
NOTE To collapse all tubes and only view the samples, right-click a tube or sample and select Collapse
All. To expand all tubes, right-click a sample and select Expand All.
Samples Collapsed
Samples Expanded
C-22 C44966AB
3 Right-click each sample and select Edit Sample ID or double-click Sample ID box to input the
Sample ID information.
[Standard Experiment mode]:
[Standard Experiment]: Skip to Step 6.
[Panel Experiment mode]:
NOTE Ensure the Sample ID information matches with the barcode label if the tube has a barcode label.
Barcode labels are not required on sample tube. Refer to Barcode Labels in CHAPTER 1, System
Overview.
C44966AB C-23
4 [Panel Experiment]: Right-click a sample then select Property. The Sample Property window
appears.
C-24 C44966AB
5 [Panel Experiment]: Set the sample properties for each sample.

a. Enter the sample type, collection time, submission time, submitted by, and test information
in the Sample Information tab of the Sample Property window.
b. Select the Patient Information tab.
C44966AB C-25
c. Enter the patient name, patient type, patient ID, gender, D.O.B, age, charge type,
department, ward, bed number, clinical diagnosis, and attending doctor in the Patient
Information tab of the Sample Property window.
d. If desired, select File > Hide Personal and Sensitive Information to encrypt the submitted by,
reviewed by, test information, the patient name, patient type, patient ID, D.O.B, charge
type, ward, bed number, clinical diagnosis, and attending doctor.
C-26 C44966AB
e. The following message appears The personal and sensitive information in the current
experiment will be replaced by “***”, are you sure you want to continue? Select Yes.
NOTE This encryption can not be retracted. Select No to cancel the encryption.
If you want to encrypt the personal and sensitive information only in the exported report, select
Hide Personal and Sensitive Information from the Batch Export Report window. Refer to
Batch Exporting Reports in CHAPTER 6, Data Acquisition and Sample Analysis.
C44966AB C-27
6 Right-click on a sample tube then select Property. The Tube Property window appears.
C-28 C44966AB
7 Set the tube properties for each tube. Enter the name, sample ID, and any desired remarks in
the Basic Information tab of the Tube Property window.
[Standard Experiment mode]
NOTE Sample ID is inactive in the Tube Property window if you are creating a new panel experiment. In
panel experiments, Sample ID is set in the Sample Property window. Refer to Steps 3.
8 Select . The Carrier window appears.
C44966AB C-29
9 Select . The Add Plate window appears.
NOTE You can add up to 1,152 plates.
C-30 C44966AB
10 Select the Plate Type and sampling sequence from the Add Plate window then select OK. The
plate image appears on the Carrier window.
C44966AB C-31
1. : Used to add a new plate.
2. : Used to delete the current plate.

3. Plate number drop-down menu: Used to toggle between plates.
4. Plate Type drop-down menu: Used to change plate types. Plate types include: 96-well flat-bottom
plate, 96-well U-bottom plate, and 96-well V-bottom plate.
5. Sampling sequence drop-down menu: Used to change the plate sampling sequence. Sampling
sequences include horizontal left-to-right, horizontal alternating left-to-right and right-to-left,
vertical top-to-bottom, and vertical alternating top-to-bottom and bottom-to-top.
6. : Used to set well locations in the plate.
7. : Used to edit the backflush setting.
NOTE The default backflush setting is 3 seconds. You can choose to set different backflush
settings for each well or press Control then select multiple wells and set backflush for all
selected wells.
8. : Used to edit the mix setting. Mix settings can be changed to an interval of X minutes, or
an interval of X wells. Mix duration and intensity can also be edited.
NOTE Mix settings vary depending on the sample injection mode selected. Single Tube Mode
does not have mix settings available. Carousel Mode only has mix duration available. Plate
Mode provides mix settings for mix duration, intensity, interval wells, and interval time.
NOTE The default mix setting is 3 seconds with an intensity of medium-high. You can choose
to set different mix time for each well or press Control then select multiple wells and set mix
time for all selected wells.
NOTE If the mix intensity is set to high, ensure the sample volume does not exceed 100 μL.
9. : Used to clear the tube location.
10. : Used to set auto acquisition.
NOTE A number displays in the bottom right corner of each well set for auto acquisition. For
example: .
11. : Used to cancel auto acquisition.
NOTE If a well is no longer set for auto acquisition, there is no number in the well location. For
example: .
11 Select the Carrier number and the sample order from the dropdowns located at the top of the
Carrier window.
C-32 C44966AB
12 Select . The Set Location window appears.
Plate Mode
13 Select the samples that should be added to the plate.

NOTE can be used to select all samples. can be used to deselect all samples.
14 Select the tube location in the plate picture.

NOTE If more than one tube is selected, the samples will automatically group together either vertically
or horizontally depending on the sample order selected. To set individual well locations in separate
locations (not grouped), select one tube at a time.
15 Select OK.
NOTE To clear a tube or well location, select the checkbox next to the tube to be cleared then select
Clear Location.
16 Change the backflush and mix setting for each well as needed.
Sampling and Collecting Data

Refer to Sampling and Collecting Data in CHAPTER 6, Data Acquisition and Sample Analysis.
C44966AB C-33
Using Heat Maps
Using Heat Maps
IMPORTANT Heat maps can only be created using a standard experiment in plate mode. This function is
not available:
• On DxFLEX instruments without the Autoloader.
• In panel experiments.
• In Carousel Mode.
• In Single Tube Mode.
Creating a Heat Map
1 Select from the Tube management controls (refer to Test Tubes in CHAPTER 2, Using the
CytExpert for DxFLEX Software). The Heat Map window appears.
C-34 C44966AB
Using Heat Maps C
2 Select from the Heat Map window. The New Heat Map window appears.
NOTE You must have at least one plate that contains data in at least two wells to create a new heat map.
NOTE Select Display Value to display the value within the heat map well on the Heat Map window.
Display Value is only selectable when using a single parameter.
3 Enter the heat map name, select the Display Name checkbox if you want the name to display in
the heat map view, and select the heat map data set from the Plate drop-down menu.
NOTE The plate drop-down menu only displays plates that contain data.
C44966AB C-35
Using Heat Maps
4 Select to add a parameter item to the Parameter section of the New Heat Map
window.
NOTE A maximum of 6 parameters can be added to a single heat map.
NOTE Select a parameter then select to delete a parameter. Select Yes when the following
message appears.
5 Change the Parameter elements in the Parameter section of the New Heat Map window.
a. Change the Label name if needed.
b. Select from the Statistic Expression section of the Parameter list. The Expression
window appears.
c. Enter the desired expression for the selected parameter then select OK. The Actual Range
displays in the Parameter section of the Heat Map window.
C-36 C44966AB
Using Heat Maps C
d. Select the Use Custom Ranges checkbox if needed and enter the Min and Max ranges.
NOTE The Actual Range displays when the statistics parameter can be calculated.
6 Select any wells that should be excluded from the heat map in the Wells section of the New Heat
Map window then select .
NOTE designates that a well is included. All wells are included by default.
NOTE designates that a well is excluded. Select any wells that should be included and select
.
C44966AB C-37
Using Heat Maps
7 Edit the color elements in the Color section of the New Heat Map window.
a. Select a color from the base color drop-down menu.
b. Select the number of color bands desired from the Bands drop-down menu. The window
refreshes to display the appropriate number of bands.
NOTE You can choose between 2 and 10 color bands.
C-38 C44966AB
Using Heat Maps C
c. Select Percentile to assign colors based on a percentage range.
NOTE If Use Custom Range is selected, the percentile is calculated according to “Min” and “Max”.
If Use Custom Range is not selected, the percentile is calculated according to “Actual Range”.
Or
Select Fixed Range to assign colors based on a fixed range specified by the user. The heat
map is created directly based on the result of the expression. The color of the heat map
displays according to the legend range.
NOTE Fixed Range can only be used with one parameter.
8 Select OK. The New Heat Map window closes to display the Heat Map window.
Heat Map with 6 Parameters
NOTE When viewing more than one parameter, the parameter location relative to the pie chart is visible
in the top, right corner of the Heat Map screen.
C44966AB C-39
Using Heat Maps
Heat Map with 1 Parameter
Refreshing a Heat Map

When data displayed in a heat map is no longer current, the symbol appears next to the heat
map name and the message Analysis changed. Please refresh. appears on the Heat Map window.
Select from the heat map toolbar to refresh the analysis.
C-40 C44966AB
Using Heat Maps C
Modifying Existing Heat Map Settings
Select from the heat map toolbar to modify existing heat map settings. The Modify Heat Map
Settings window appears.
C44966AB C-41
Daily Clean [With Autoloader - Plate Mode]
Deleting an Existing Heat Map

To delete a single heat map, select the heat map to be deleted from the list of heat maps in the Heat
Map window then select from the heat map toolbar.
To delete multiple heat maps, select from the heat map toolbar. The Delete Multiple Heat
Maps window appears.
Select the heat maps to be deleted then select OK.
NOTE The Select All checkbox allows you to delete all of the heat maps listed.
Exporting a Heat Map

Heat maps can be exported as a graphics file (.bmp or .emf) or to a clipboard (.bmp).
To export a heat map as a graphics file, select from the heat map toolbar.
To export a heat map to a clipboard, select from the heat map toolbar.
Daily Clean should be performed during instrument startup and instrument shutdown to clean the
sample line.
After sampling an excessively large sample or a sample that can easily clog the sample probe, it is
recommended to perform the Daily Clean procedure.
C-42 C44966AB
Daily Clean [With Autoloader - Plate Mode] C
For instructions on performing Daily Clean with the Autoloader using the Single Tube Mode, or
Carousel Mode, refer to Daily Clean [With Autoloader - Single Tube and Carousel Mode] in
3 Select Daily Clean in the Cytometer menu. The Daily Clean window appears.
C44966AB C-43
IMPORTANT You must select at least one cleaning solution well and one water well.
4 Follow the on screen software prompts and select the desired wells for cleaning agent and
deionized water.
The plate automatically sets the default well locations for the cleaning agent and deionized
water wells as shown in the figure above. indicates cleaning agent and indicates
deionized water.
NOTE To deselect wells, select the desired well and select Set As Empty.
NOTE To select new cleaning agent well locations, select the desired wells and select Cleaning Agent.
To select new deionized water well locations, select the desired wells and select Deionized Water.
5 Select Start to start the cleaning procedure. The message Please confirm that the correct plate is
placed properly and press OK appears. Select OK.
NOTE Select Stop at any time to stop the daily clean process.
6 When daily clean is complete, select Close.
C-44 C44966AB
Calibrating the Sample Flow Rate in Plate Mode C
Calibrating the Sample Flow Rate in Plate Mode
Calibrate the sample flow rate:
• After replacing the sample peristaltic pump tubing.

• If a precise volumetric measurement is required.
The accuracy of concentration calculations can be affected by the sample flow rate.
Refer to Calibrating the Sample Flow Rate in CHAPTER 12, Replacement/Adjustment Procedures for
instructions on calibrating the Sample Flow Rate in DxFLEX [without Autoloader] or DxFLEX [with
Autoloader-Single Tube Mode] and DxFLEX [with Autoloader-Carousel Mode].
1 Verify that the instrument is in the initialized state.
2 Select Calibrate Sample Flow Rate in the Cytometer menu.
C44966AB C-45
3 Select the flow rate to be calibrated.

If you want to calibrate all flow rates at once, fast calibration is recommended. The rate selected
in the Calibrate Sample Flow Rate window overrides the rate selected in the Acquisition
window.
4 Prepare one sample tube with 1 mL of clean deionized water then use a calibrated analytical
balance to measure the weight of the prepared sample tube. Record the weight and enter it into
the software.
With Autoloader (Plate Mode)
NOTE DxFLEX [With Autoloader]: Select in the Calibrate Sample Flow Rate window
to set the well location and the Carrier Type.
Input weight: 0-100
Collection time (score): 1-30
5 Select Next and put the well plate in the sample loading position (see Figure 1.13).
C-46 C44966AB
Calibrating the Sample Flow Rate in Plate Mode C
6 Select Initialize to start the sample run.
With Autoloader (Plate Mode)
7 Select Run to begin acquiring the sample.

DxFLEX [With Autoloader - Plate Mode]: The following system message appears Please confirm
that the correct plate is placed properly and press OK. Select OK.
C44966AB C-47
8 Wait for the sample run to finish, remove the sample tube, and use the analytical balance to
measure the weight and record the value.
9 Select Next to determine if the results fall within the acceptable range.
If the results fall within the acceptable range, the current setting is kept.
If deviation occurs, the setting is automatically corrected.
NOTE Select [According to this value calibrate other flow rate] to apply the coefficient directly to
skip calibrating the Medium and Slow flow rate.
C-48 C44966AB
Calibrating the Carrier in Plate Mode C
Calibrating the Carrier in Plate Mode
Use the following procedure to calibrate the carrier position:
• Upon installation
• After a new carrier type, carousel type, or plate type is defined for first use
• After changing plate manufacturers of the same previously calibrated plate
• When optimizing carrier performance
Refer to Calibrating the Carrier [DxFLEX With Autoloader] in CHAPTER 12,

Replacement/Adjustment Procedures for instructions on calibrating the Carrier in Single Tube or
Carousel Mode.
1 Ensure the sample injection control mode is set to Plate Mode. Refer to Selecting the Proper
Sample Injection Mode [with Autoloader] in CHAPTER 4, Daily Startup.
2 Select Advanced > Calibrate Carrier. The Calibrate Carrier window appears.
3 Select the plate type and place the plate adapter and the plate inside the autoloader.
C44966AB C-49
4 Select Calibrate. The message Please confirm that the correct carrier is placed properly and press OK.
appears. Select OK.
CAUTION
Risk of instrument damage. Do not crash the probe into the bottom of the plate as
this will cause irreparable damage to the probe. Move the probe one step at a time
when lowering the probe on the Z-axis toward the bottom of the plate. You will
hear a click when the probe makes contact with the bottom of the well. Do not
move the probe any lower after you hear the click. Move the probe back up three
steps after you hear the click, this is the correct position for the Z-axis of the
probe.
5 Select and or and to adjust the sample probe positions in

the X-, Y-, and Z-axes.
Ensure the sample probe is centered and touches the bottom of the well.
NOTE The sample probe should just make contact with the bottom of the well.
NOTE The X-axis arrows moves the sample probe well position left and right. The Y-axis arrows moves
the sample probe well position forward and back. The Z-axis arrows moves the sample probe up and
down.
6 Select Test to verify the sample probe position.

Listen for the click to ensure the probe has made contact with the bottom of the well. Readjust
the sample probe position if necessary.
C-50 C44966AB
7 Select Next to move to the next well.
8 Repeat Steps 5-7 for the remaining three corner wells.
C44966AB C-51
9 Select additional plate types to be calibrated on the Calibration completed window, if

necessary.
Place the corresponding plate on the plate holder and repeat Steps 5-8.
10 Select Finish to save the settings.
C-52 C44966AB
11 Select Close to quit the program.
12 Reinstall the front cover. Refer to Front Cover Removal in CHAPTER 12,
C44966AB C-53
C-54 C44966AB
APPENDIX D
Good Practices for Cyber Security
Overview
The following procedures are the only validated and approved recommendations for cyber privacy
and security. Contact your IT professional for assistance.
• Changes below require Administrator Level Windows access.

• Before connecting to an external device such as a hard drive, USB, or DVD/CD, verify it does not
have a virus or malware.
• Close all unnecessary applications while operating the CytExpert software.
CAUTION
System integrity could be compromised and operational failures could occur if:
• You introduce software that is not authorized by Beckman Coulter into your
computer. Only operate your system’s computer with software authorized by
Beckman Coulter.
• You apply domain policies which alter the default configuration.
• You alter the system in a manner other than the approved changes outlined
below.

• Drive Encryption
• Protection from Malware Software
• Operating System Updates
• System Hardening
• Remote Access
Drive Encryption
Your Windows 10 system is equipped with BitLocker. Use BitLocker disk encryption software to
prevent unauthorized access to your hard drive, refer to BitLocker Keys. For instructions on
BitLocker Encryption or BitLocker Decryption, see Enabling BitLocker or BitLocker Decryption.
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Drive Encryption
IMPORTANT BitLocker is the only approved encryption method.
BitLocker Keys
CAUTION
Risk of data loss. Store your BitLocker key in a safe place. For instructions on
saving your BitLocker key, refer to Backing Up Your Recovery Key. If you lose your
BitLocker Key, you will lose access to your entire hard drive including all of your
data. Beckman Coulter is not responsible for security keys and will not be able to
recover your data off of your hard drive. Work with your IT department in creating
and storing the keys in a safe place.
Changing Encryption Strength Level for BitLocker

BitLocker defaults to XTS-AES 128-bit encryption method. To alter the encryption, follow the steps
below before enabling BitLocker. Changes to the encryption level will only impact the encryption
of new volumes. Work with your IT department to change the Encryption Strength Level.
Changing the encryption strength for BitLocker
1 Type gpedit.msc from the Windows search bar to locate the gpedit Control Panel, see Figure D.1.
Figure D.1 gpedit Search
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Drive Encryption D
2 Click gpedit.msc. The Local Group Policy Editor window appears.
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Drive Encryption
3 Navigate to Computer Configuration > Administrative Templates > Window Components >
BitLocker Drive Encryption.
4 Double-click “Choose drive encryption method and cipher strength (Windows 10.....)”. Refer to
Figure D.2.
Figure D.2 Choosing Drive Encryption Method and Cipher Strength
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Drive Encryption D
5 Select Enabled and use the drop-down arrows to select the encryption method for the operating
system drive, fixed data drive, and for the removable data drives, see Figure D.3.
Figure D.3 Choosing the Encryption Level
NOTE The default is set to XTS-AES 128-bit.
6 Select Apply.
7 Select OK and close all windows.
8 Restart the computer and proceed to Enabling BitLocker.
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Drive Encryption
Enabling BitLocker
1 Type Manage BitLocker from the Windows search bar to locate the Manage BitLocker Control
Panel.
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Drive Encryption D
2 Double-click Manage BitLocker to open the BitLocker Drive Encryption window.
3 Select Turn on BitLocker from the Control Panel screen.
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Drive Encryption
4 BitLocker verifies that your PC meets the requirements. Refer to Figure D.4.
Figure D.4 Checking PC Configurations
5 Select Print the recovery key to back up the recovery key. For instructions, refer to Backing Up
Your Recovery Key.
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Drive Encryption D
6 Select “Encrypt used disk space only (faster and best for new PCs and drives)”, then select Next.
7 Select “New encryption mode (best for fixed drives on this device)”, then select Next.
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Drive Encryption
IMPORTANT The system check ensures that BitLocker can read the recovery and encryption keys correctly
before encrypting the drive. This check may take some time, but it is recommended to ensure that your
selected unlock method works without requiring the recovery key.
8 Select “Run BitLocker system check”, then select Start encrypting.
NOTE BitLocker restarts your computer before encrypting.
9 Select Restart now to start the BitLocker Encryption.
10 Enter your password in the Password screen.

NOTE After logging in you will see a Window that says Encrypting in Progress.
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Drive Encryption D
11 Open the Manage BitLocker Control Panel from the Windows search bar.
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Drive Encryption
12 Under Operating System Drive, you will see the BitLocker is Encrypting the drive. See
Figure D.5.
Figure D.5 BitLocker Encrypting the Drive
13 Once the BitLocker has encrypted the drive, the BitLocker will be on. See Figure D.6.
Figure D.6 BitLocker Successfully On
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Drive Encryption D
Backing Up Your Recovery Key

IMPORTANT The Back Up Your Recovery Key to your Microsoft Account is not supported at this time.
Panel, see below.
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Drive Encryption
3 Select “Backup your recovery key”. The following window appears.
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Drive Encryption D
IMPORTANT The Recovery Key can only be saved to a USB or external drive. You cannot save to a location
on your PC.
4 Select a destination to save the Recovery Key.

• Save to a file: Select “Save to a file”. The following screen appears.
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Drive Encryption
• Print the recovery key: Select “Print the recovery key”. The following screen appears.
1. Select “Microsoft Print to PDF”, and then select Print.
2. Select the location that you want the file saved. In the File name, type BitLocker Key and
in the Save as type, ensure that it is listed as.pdf.
5 Select Save then select Finish.
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Drive Encryption D
BitLocker Decryption
IMPORTANT Turning off the BitLocker feature decrypts all the data on the drive. Decrypting your operating
system means that you have removed BitLocker from your system. The BitLocker key will change each
time BitLocker is re-enabled. Only the latest BitLocker key will work. Previous BitLocker keys will be
invalid and will not unlock the system. Please note this is a time-consuming process. However, you can
also suspend BitLocker instead of decrypting the drive, see Suspending BitLocker Drive Encryption.
Turning off BitLocker
Panel.
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Drive Encryption
2 Double-click Manage BitLocker to open the BitLocker Drive Encryption.
3 Select Turn off BitLocker.
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Drive Encryption D
The following message appears.
IMPORTANT Drive decryption might take a long time, but you can keep using your PC during the decryption
process.
4 Select Turn Off BitLocker. The BitLocker Drive Decryption screen displays.
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Drive Encryption
After the decryption has completed, the following screen displays.
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Drive Encryption D
Suspending BitLocker Drive Encryption

IMPORTANT Suspending BitLocker Drive Encryption is a temporary method for removing BitLocker
protection without decrypting the Windows drive. Use Suspend BitLocker if you need to update the BIOS
firmware or Windows boot loader/startup files that requires access to the drive in a pre-boot environment.
This will help prevent BitLocker from locking the drive and may avoid a lengthy decryption process.
Suspending BitLocker
Panel.
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Drive Encryption
3 Select Suspend Protection.
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Drive Encryption D
A confirmation message “Do you want to suspend BitLocker protection?” displays.
IMPORTANT If you choose to suspend BitLocker, your data will not be protected.
4 If you want to proceed, select Yes and the following popup information will display.
If you do not want to proceed, select No.

NOTE There are certain situations where you may want to suspend BitLocker such as updating your PC’s
firmware, hardware, or operating system. If you forget to resume BitLocker after these updates are
completed, BitLocker will resume automatically the next time you restart your PC.
Restarting BitLocker Protection
1 Select Resume Protection.
Recovering Your BitLocker Key

There may be situations where the BitLocker Key is requested from your computer, such as:
• Your drive is placed onto another computer

• Computer requests your recovery key, it may be due to one of the following but is not limited to:
— Hardware changes
— Firmware changes
— Other Windows updates
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Drive Encryption
BitLocker Recovery
1 BitLocker Recovery window will be displayed upon powering on your system.
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Protection from Malware Software D
2 Enter your key from your saved text file or printed copy, then select Enter.
3 Windows will reboot your computer.
CAUTION
The instrument workstation is vulnerable to malware, viruses, and unauthorized
access if the DxFLEX PC is connected directly to the laboratory network without
installation of all approved OS and malware patches.
IMPORTANT Only the version of the McAfee® Application Control (Version 8.3) has been validated to work
with the DxFLEX instrument.
For more information on Configuration of McAfee® Application Control, refer to McAfee®

Application Control Guide.
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Installing McAfee Application Control
IMPORTANT The McAfee® Application Control Software (version 8.3) should be purchased separately from
McAfee. Download the McAfee® Application Control Software, access McAfee website at
www.mcafee.com, or find it from the CytExpert software disk provided to you.
1 Unzip the McAfee Application Control file SOLIDCORxxx_xxx_WIN.zip
2 Right-click Setup-win-8-10-2010-amd64-xxx.exe, and select Run as administrator. The McAfee

Installer opens.
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3 Select Next. The License Agreement displays.
Read the license agreement and select I accept the terms in the license agreement, then select
Next.
4 Enter the user name and the serial number.
If you select Install without license key, follow the procedures below to add the license key after
the installation.
C44966AB D-27
a. Right-click on your desk top, and select Run as administrator. The following window
displays.
b. Enter cd C:\Program Files\McAfee\Solidcore.
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c. Enter the license key in the console.
The license key is added.
5 Select Next.
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6 Select the destination folder and select Next. The system starts to install McAfee Solidifier.
7 Select Finish.
Enabling McAfee Application Control

The enabled McAfee® Application Control Software prevents your system from executing
unauthorized applications on your instrument workstation.
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IMPORTANT Ensure all the necessary applications have been installed prior to enabling the
McAfee®Application Control.
1 Right-click on your desk top, and select Run as administrator. The following window
displays.
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2 Enter cd C:\Program Files\McAfee\Solidcore.
3 Enter sadmin so in the console to scan the system and create a white list. The solidification
process takes about 10 minutes depending on your system size and CPU performance.
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4 Enter sadmin enable to enable the McAfee Application Control.
IMPORTANT Beckman Coulter recommends setting a password to manage the authority of

enabling/disabling the McAfee Application Control.
5 Enter sadmin passwd to set a password to lock the McAfee Application Control authority.
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6 Restart the system.
Exporting the Certificate

Follow procedures below to export the certificate of CytExpert software into your PC.
1 Right-click on the application CytExpert for xxxx Setup, and select Properties. The following
window displays.
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2 Select the Digital Signatures tab. Then select the signature and select Details to view the
signature information.
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The Digital Signature Details window displays.
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3 Select View Certificate.
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4 Select the Details tab and then select Copy to File.
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The Certificate Export Wizard window appears.
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5 Select Next to export the certificate.
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6 Select Base-64-encoded X.509 and then select Next.
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7 Select a destination to save the certificate and then select Save.
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8 Select Next.
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9 Select Finish when the success message displays.
Importing BEC Certificate

Follow the procedures below to add the certificate of CytExpert software into the McAfee Certificate
list. All the certified applications will be regarded as trusted and authorized by the McAfee
Application Control.
IMPORTANT McAfee Application Control recognize only X.509 certificates.
1 Copy the certificate BEC.cer from the CytExpert software disk into your instrument PC.
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displays.
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4 Enter sadmin cert add C:\BEC.cer to add the certificate.
Switching McAfee Application Control to Update Mode

The update mode allows the McAfee Application Control to scan the instrument workstation and
add the newly-installed application into the white list automatically. Switch the McAfee
Application Control to the update mode when you execute scheduled or emergency changes on
your instrument workstation.
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displays.
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3 Enter sadmin bu to switch the McAfee Application Control to Update Mode.
4 Execute scheduled or emergency changes, patch installations, or software updates for your
instrument workstation.
5 Enter sadmin eu to exit the Update Mode.
Disabling McAfee Application Control

IMPORTANT Disabling McAfee Application Control is a temporary method in special cases, for example,
Windows or firmware changes. This will help stop McAfee Application Control from blocking unauthorized
applications. Beckman Coulter recommends enabling the McAfee Application Control in most cases.
All the applications installed in this mode will not be included in the McAfee white list.
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displays.
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3 Enter sadmin disable in the console.
4 Select Enter.
5 Restart the System.
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6 Enter sadmin status to view the status of McAfee Application Control.
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WARNING
Risk of software incompatibility. Only operating system updates posted to the
Beckman Coulter website should be installed on the instrument controller PC. Do
not install patches from any other source.
Install the update on one instrument and verify functionality before installing on
other instruments.
Enabling System Protection
1 In the Start Menu, type Restore to locate the Create a Restore Point.
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Operating System Updates D
2 Double-click Create a Restore Point to open the System Properties window.
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3 Select Configure.
The following window appears.
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4 Select Turn on system protection, then select Apply.
5 Close the System Properties window by selecting OK.
Creating a Restore Point

A restore point is used to remove all software and update installations that occurred after it was
created. If installing an update on your PC causes any loss of functionality, the restore point may be
used to place the system back into the operable state present before the update installation. In all
cases, a system restore with any restore point would not cause loss of any data (i.e. LMD files) on the
system.
CAUTION
Risk of unexpected software modification. To ensure a system recovery point is
available that is indicative of the most recent software configuration, the user
should create a system restore point immediately prior to running the operating
system update installer. Otherwise, a user would need to utilize the most recent
automatically created restore point which may be up to 72 hours old and not
include all recent software modifications.
1 In the Start Menu, type Restore to locate the Create a Restore Point.
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3 Type a name for the restore point and select Create.
4 Wait for the restore point creation to complete and then select Close.
5 Close the System Properties window by selecting OK.
Downloading Operating System Updates

In order to download and install operating system updates, you will need to access the Beckman
Coulter website at www.beckmancoulter.com.
IMPORTANT Account registration is required for new users.
1 Go to www.beckmancoulter.com.
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2 Using the menu bar in the in the upper right hand corner of the screen, hover over Login, then scroll
down to select Software Downloads.
3 To search by product:
a. Select Research & Discovery from the Market Segment drop-down menu.
b. Select Clinical Flow Cytometry from the Product Line. drop-down menu.
c. Select Instruments from the Product Series drop-down menu.
d. Select DxFLEX from the Product drop-down menu.
4 Select Search.
IMPORTANT Print out the Release Notes for each update as they will not be available when the next update
is released. The Component Name section of the release notes identifies the Microsoft updates that are
included in the download. This information may be requested by your IT team and will be needed to
recover from a failed update.
5 Each operating system update is cumulative and only the most recent validated set of updates
will be available for download. Select the first file in the results to view the release notes and
then select .
If more than one update is listed in the search results, select , and repeat this step for
each update.
If the operating system updates cannot be not downloaded directly to the instrument
controller system, transfer the downloaded files via an encrypted external media or secured
network drive to the instrument controller PC.
Notification Options
To receive notification when a new operating system update is validated and available for
download, you must sign up for an account on www.beckmancoulter.com and follow the procedure
for setting up subscriptions.
Sign up for subscriptions as outlined below:

• Select the Document Language consistent with your IFU language.
• Select Research & Discovery from Market Segment.
• Select Clinical Flow Cytometry from Product Line.
• Select Instruments from Product Series.
• Select DxFLEX from Product.
If you do not sign up for product notifications, you will have to check periodically for newly released
operating system updates, see Downloading Operating System Updates.
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Installing the Operating System Updates
CAUTION
Risk of software incompatibility. Beckman Coulter can only support operating
system updates on unmodified systems. Application of domain policies or other
configuration changes may impact compatibility.
IMPORTANT If more than one update is available for your product, install the updates in the order indicated
by the sequence number as outlined in the file naming convention below.
RSU- <operating system>-<product name>-<Year><Month>_<Sequence #>
Example: RSU-W10LTSC-DxFLEX-2020Jun_1_0
1 Ensure you have a recent copy of your BitLocker key, refer to Backing Up Your Recovery Key.
2 Login to Windows as a user with Administrator access.
3 Close all program and save open files.
4 Power down the instrument.
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5 Open the first operating system update file (For example: RSU-W10LTSC-DxFLEX-2020Jun_1_0)
on the Instrument Controller PC by double-clicking on the file. If the Microsoft Windows
Defender SmartScreen prompt appears, select More Info and Run Anyway to launch the installer.
This process may take several minutes. See Figure D.7.
Figure D.7 Security Update Setup Installing
NOTE The security update installer will automatically detect and install only the necessary updates.
6 Once the operating system update setup has completed, select OK to restart the system and
complete the update process. See Figure D.8 and Figure D.9.
Figure D.8 Security Update Setup Successfully Installed
Figure D.9 Windows Shutdown
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7 Wait while the system completes the installation of the security updates. This process may take
several minutes and will return you to the Windows login screen. See Figure D.10.
Figure D.10 Working on Completing Security Updates
8 Install any additional downloaded operating system updates (sequence numbers above 1) by
repeating steps 4 to 6.
9 Verify functionality of instrument software before installing operating system updates on any
other system. If functionality is impaired, see Recovering from Failed Operating System
Updates.
Recovering from Failed Operating System Updates

These steps below should only be followed if the instrument software or operating system
functionality is impaired after an operating system update was applied.
1 Power down the instrument and close all programs. Do not turn your workstation off.
2 In the start menu, type view update and select View your Update History.
3 Select Uninstall Updates.
4 Highlight the update listed under Microsoft Windows that matches the cumulative security
update KB number in the release notes. Refer to Component Name section of the Release Notes,
see Step 5, Downloading Operating System Updates.
5 Select Uninstall and select Yes when prompted to verify the uninstall. This process may take
several minutes to complete.
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6 When prompted to restart, select Restart Now.
7 Once the PC has been rebooted, login and verify Microsoft update has been removed by
repeating Steps 2 and 3. Verify the KB selected in Step 4 is no longer listed.
8 Power on the instrument and verify restored software functionality.

• If software functionality has been restored, continue using your instrument without the
update installed. Contact us for troubleshooting the update incompatibility. Do not
proceed to Step 9.
• If software functionality has not been restored, continue to Step 9.
9 In Start Menu, type Restore and select Create a Restore Point.
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11 Select System Restore. The following window appears.
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12 Select Next. The following window appears.
WARNING
Risk of software changes. All software installations and operating configuration
system changes that occurred after the selected restore point timestamp will be
removed during the restore process.
13 Select the most recent Restore Point and select Next. The following window appears.
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14 Select Finish. The confirm message appears.
15 Select Yes. Wait while the system restores. This process takes about several minutes to return
to the Windows login screen.
IMPORTANT Operating system updates removed during a restore operation will remain listed on the
windows update history screen.
16 Log in to the system again and test the functionality of software.
17 Contact us for assistance in troubleshooting the update incompatibility or for assistance

restoring functionality.
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System Hardening D
System Hardening
Enabling Automatic Log-Off

IMPORTANT Automatic log-ff can protect your instrument PC from unauthorized operation when you leave
the PC temporarily. Follow the steps below to enable the Screen Saver protection.
Enabling Screen Saver Protection
1 Type Edit group policy from the Windows search bar to locate the Edit group policy editor.
2 Double-click Edit group policy editor to open the Local Group Policy Editor window.
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System Hardening
3 Navigate to User Configuration > Administrative Templates > Control Panel > Personalization.
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System Hardening D
4 Select Enable screen saver. The following window appears.
5 Select Enabled and Apply.
6 Select OK to close the Enable screen saver window.
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System Hardening
7 Select Password protect the screen saver from the Local Group Policy Editor window.
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System Hardening D
The Password Screen saver window appears.
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System Hardening
9 Select OK to close the window.
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System Hardening D
10 Select Screen saver timeout from the Local Group Policy Editor window.
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System Hardening
The Screen saver timeout window appears.
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System Hardening D
Setting Lock Screen
1 Type Lock screen settings from the Windows search bar to locate Screen saver settings.
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System Hardening
The Settings window appears.
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System Hardening D
2 Scroll down to the bottom and select Screen saver settings. The Screen Saver Settings window
appears.
3 Select Apply.
Setting Account Lockout Policy
1 Type Local Security Policy from the Windows search bar to locate the Local Security Policy.
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System Hardening
2 Double-click Local Security Policy to open the Local Security Policy window.
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System Hardening D
3 Navigate to Security Setting > Account Policies >Account Lockout Policy.
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System Hardening
4 Customize the account lockout policy.

a. Double-click on Account lockout threshold to set the account lockout threshold.
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System Hardening D
b. Double-click on Account lockout duration to set the account lockout duration.
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System Hardening
c. Double-click on Reset account lockout counter after to reset the account lockout counter.
NOTE The Account lockout duration and Reset account lockout counter after settings only have
meaning when an Account lockout threshold is specified.
Setting Password Policy
1 Type Local Security Policy from the Windows search bar to locate the Local Security Policy.
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System Hardening D
2 Double-click Local Security Policy to open the Local Security Policy window.
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System Hardening
3 Navigate to Security Setting > Account Policies >Password Policy.
4 Customize the password policy.

• Enforce password history
• Maximum password age
• Minimum password age
• Minimum password length
• Minimum password length
• Password must meet complexity requirements
• Store passwords using reversible encryption
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System Hardening D
Disabling Remote Access
1 Type Control Panel from the Windows search bar to locate the Control Panel app.
2 Double-click Control Panel to open the Control Panel window.
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System Hardening
3 Type Security from the search bar to locate Allow remote access to your computer.
4 Select Allow remote access to your computer. The System Properties window appears.
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System Hardening D
5 Select Don’t allow remote connections to this computer from the Remote tab.
Disabling Auto Play
1 Type Auto Play settings from the Windows search bar to locate the Auto Play settings.
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System Hardening
2 Select Auto Play settings. The Settings window appears.
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System Hardening D
3 Select Off to turn off Auto Play.
4 Close the window.
Disabling Unnecessary Services

It is recommended to disable all the unnecessary services while operating the CytExpert software.
Refer to Table D.1.
Table D.1 List of Unnecessary Services
Bluetooth Support Service Microsoft FTP Service (If Windows Media Player Network
available) Sharing Service
Geo-location Service Microsoft iSCSI Initiator Service Windows Mobile Hotspot Service
Infrared monitor service Remote Procedure Call (RPC) WinHTTP Web Proxy Auto-
Locator Discovery Service
Internet Connection Sharing Routing and Remote Access Xbox Live Game Save
Link-Layer Topology Discovery SSDP Discovery Xbox Live Networking Service
Mapper
LxssManager (If available) UPnP Device Host
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System Hardening
1 Type Computer Management from the Windows search bar to locate the Computer Management
App.
2 Select Computer Management.
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System Hardening D
3 Navigate to Services and Applications > Services.
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System Hardening
4 Select the service you want to disable.
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System Hardening D
5 Select Disabled from the Startup type dropdown.
6 Select Apply.
Disabling Unauthorized Applications
1 Type Edit group policy from the Windows search bar to locate the Edit group policy editor control
panel.
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System Hardening
3 Navigate to Computer Configuration > Administrative Templates > Windows Components > Store.
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System Hardening D
4 Select Disable all apps from Windows Store.
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System Hardening
7 Disable the following applications. Refer to Steps 4-6.

• Turn off Automatic Download and Install of updates.
• Turn off the offer to update to the latest version of Windows.
• Turn off the Store application.
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System Hardening D
Enabling Installation Restriction
1 Type Edit group policy from the Windows search bar to locate the Edit group policy editor control
panel.
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System Hardening
3 Navigate to Computer Configuration > Administrative Templates > Windows Components >
Windows Installer.
4 Select Allow user control over installs.
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System Hardening D
5 Select Disabled and Apply.
7 Disable install with elevated privileges. Refer to Steps 4-6.
Enabling Firewall Defender
1 Type Windows Defender Firewall from the Windows search bar to locate the Control Panel.
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System Hardening
2 Select Turn Windows Defender Firewall on or off.
3 Select Turn on Windows Defender Firewall for both Private network and Public network settings.
4 Select OK.
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System Hardening D
Enabling Network Time Protocol

To ensure the accuracy of date and time information included in logs, follow the instructions below
to enable the Network Time Protocol.
1 Type Change the date and time from the Windows search bar.
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System Hardening
2 Double-click Change the date and time. The Date and Time Settings window appears.
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System Hardening D
3 Select Set time automatically to access the Date and Time window.
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System Hardening
4 Select Change settings from the Internet Time tab.
The Internet Time Settings window displays.
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Remote Access D
5 Select Synchronize with an internet time server, and select OK. The Cytometer PC is set to
automatically synchronize with the internet time server.
Remote Access
IMPORTANT Use BeckmanConnect for remote control over the DxFLEX instrument. For instructions on
using the BeckmanConnect, refer to the BeckmanConnect Installation and Activation Instructions.
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Remote Access
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APPENDIX E
Table of Hazardous Substances
The Hazardous Substances Names and Concentration is shown in Table E.1
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E-2
Table E.1 Table of Hazardous Substances Name and Concentration

电子电气产品号码产品名称 Product Name: 流式细胞仪 Flow Cytometer
EEP Part Number: C44326 产品型号 Product Model Number: DxFLEX
部件名称有毒有害物质或元素 Hazardous Substances Name
Component Name 铅汞镉六价铬多溴联苯多溴二苯醚
(Pb) (Hg) (Cd) (Cr6+) (PBB) (PBDE)
印刷电路板组件 Circuit Boards X O O O O O
电源组件 Power Supplies O O O O O O
计算机 Computer O O O O O O
功率调节器 Power Conditioner O O O O O O
光量传感器 Optical Sensors O O O O O O
激光 Laser O O O O O O
发动机/泵/阀门/ Motors/Pumps/Valves O O O O O O
电线 Cables X O O O O O
管路及橡胶 Tubing & Rubber O O O O O O
塑料部件 Plastic O O O O O O
连接部件 Hardware O O O O O O
包装材料 Packing Material O O O O O O
This table is prepared in accordance with the provisions of SJ/T 11364
○：表示该有毒有害物质在该部件所有均质材料中的含量均在GB/T 26572标准规定的限量要求以下
x: 表示该有毒有害物质至少在该部件的某一均质材料中的含量超出GB/T 26572标准规定的限量要求
（企业可在此处，根据实际情况对上表中打“×”的技术原因进行进一步说明)
O: Indicates that the toxic or hazardous substances contained in all of the homogenous materials for this part is below the limit requirements in GB/T 26572.
X: Indicates that the toxic or hazardous substance contained in at least one of the homogenous materials used for this part in above the limit requirement in
GB/T 26572.
C44966AB
Abbreviations
The following list is a composite of the symbols, abbreviations, acronyms, and reference
designators either used in this manual or related to the information in it. When the same
abbreviation (or reference designator) is used for more than one word (or type of component), all
meanings relevant to this manual are included, separated by semicolons.
' — foot CDRH — Center for Devices and Radiological

Health
“ — inch
CFSE — carboxyfluorescein succinmidyl ester
% — percent
cm — centimeters
°C — degrees Celsius
CSV — comma separated value
°F — degrees Fahrenheit
CV — coefficient of variation
± — plus or minus
DNA — deoxyribonucleic acid
< — less than
ECD — Energy Coupled Dye
> — greater than
EFUP — Environmentally friendly Use Period
 — less than or equal to
EMF — enhanced metafile format
μ — micron
EMR — electromagnetic radiation
μL — microliters
FAPD — Fiber Array Photo Detector
μm — micron
FCS — flow cytometry standard
A — ampere
FITC — Fluorescein isothiocyanate
AC — alternating current
FSC — forward scatter
APC — Allophycocyanin
GB — gigabyte
APC-A700 — Allophycocyanin-Alexa
Fluor™ 700 tandem dye GHz — gigahertz
APC-A7500 — Allophycocyanin-Alexa Gr Wt — gross weight

Fluor™ 750
H — humidity
APC-Cy7 — Allophycocyanin-Cyanin 7
Hz — hertz
Acq. — Acquisition
IEC — International Electrotechnical
BCI — Beckman Coulter Incorporated Commission
BMP — bitmap IVD — In vitro diagnostic
BP — band-pass filter kg — kilograms
KO — Krome Orange
C44966AB Abbreviations-1
Abbreviations
LED — light emitting diode RH — relative humidity
L — liter RoHS — Restriction of Hazardous Substances

Directive
LJ — Levey-Jennings
RPTM — real-time messaging protocol
LWH — length, width, height
S/N — serial number
m — meter
SNR — signal to noise ratio
MB — megabyte
SSC — side scatter
MFI — median fluorescence intensity
USB — universal serial bus
Mhz — megahertz
V — volts
min — minute
VA — volt-ampere
mL — milliliter
VAC — voltage alternating current
mm — millimeter
WDM — wavelength division multiplexer
mW — milliwatt
W — watts
NA — numerical aperture
NaClO — sodium hypochlorite solution
NaN3 — sodium azide
nm — nanometer
Nt Wt — net weight
PB — Pacific Blue™ dye
PC5 — Phycoerythrin-Cy™5 tandem dye
PC5.5 — Phycoerythrin-Cy™5.5 tandem dye

PC7 — Phycoerythrin-Cy™7 tandem dye
PE — Phycoerythrin
PEEK — polyether ether ketone

PerCP — Peridinin-Chlorophyll
PI — Propidium Iodide
PN — part number
QC — quality control
RAM — random access memory
rCV — robust coefficient of variation
Abbreviations-2 C44966AB
References
1. American Identification Manufacturer’s group (AIM), Uniform Symbology Specifications Code 39,
Interleaved 2 of 5, Codabar, and International Symbology Specifications Code 128. ANSI/AIM BC1,
BC2, BC3, BC4, 1995. http://www.aimusa.org
2. American National Standards Institute (ANSI) Barcode Print Quality Guidelines. X3. 182-1990
(R2000). http://www.ansi.org
3. Clinical and Laboratory Standards Institute (CLSI), Laboratory Automation: Barcodes for
Specimen Container Identification; Approved Standard. AUTO2-A. http://www.clsi.org
C44966AB References-1
References
References-2 C44966AB
Index
Symbols status bar, 2-9

’ test tubes, 2-6
define, Abbreviations-1 acquisition settings
" to configure, 6-53
define, Abbreviations-1 add
°C vertex to auto polygon gate, 6-31
define, Abbreviations-1 add page
°F report, 8-19
define, Abbreviations-1 adding
μ barcode labels (autoloader), 1-22
define, Abbreviations-1 Deep Clean solution, 12-14
μL standardization item, 5-27
define, Abbreviations-1 tubes to carousel in autoloader, 1-23
μm adding barcode labels
define, Abbreviations-1 autoloader, 1-22
± adding channels
define, Abbreviations-1 for compensation, 7-18
< adjust compensation
define, Abbreviations-1 manually, 7-12
> adjusting
define, Abbreviations-1 autogate movement, 6-32
compensation, 7-12
compensation settings, 8-8
Numerics gain, 6-54
3-of-9 threshold, 6-55
bar-code, B-5 adjusting auto gates, 6-30
adjustment
nonscheduled, 12-35
A routine, 12-2
A advanced menu, 2-17
define, Abbreviations-1 alarm
AC Fluidics module, 1-10
define, Abbreviations-1 mute, 1-10
account alarm does not sound when the waste container
unlock, 2-22 is full or the sheath fluid container is low
account policies, 2-26 and the software status display is
acoustic noise level, 1-28 red, 10-12
Acq. analysis
define, Abbreviations-1 batch samples and tubes, 8-24
acquisition screen, 2-3 screen, 8-1
collection, 2-5 to cancel, 8-23
navigation, 2-4 analysis screen, 2-10
plot area, 2-8 to open, 8-3
Index-1
Index
analyze backup
sample, 8-22 database, 10-23
samples, 8-21 manual injection mode, 4-9
tube, 8-22 band-pass filter, 1-7, Abbreviations-1
tubes, 8-21 band-pass filters, 1-6
analyzing bar-code
data, 6-60 3-of-9, B-5
APC acceptable, B-4
define, Abbreviations-1 Codabar, B-5
APC-A7500 Code 128, B-5
define, Abbreviations-1 Code 39, B-5
APC-Cy7 error rate, B-4
define, Abbreviations-1 I2-of-5, B-5
applying Interleaved 2-of-5, B-5
standardization in QC, 5-32 NW7, B-5
auto export settings, 2-47 specifications, B-1
auto gates symbologies, B-4
adjusting, 6-30 USD-1, B-5
creating, 6-30 USD-1.25, B-5
auto polygon gate USD-3, B-5
add vertex, 6-31 USD-4, B-5
auto recalculate USD-6, B-5
turn off, 6-31 use of checksum, B-7
turn on, 6-31 bar-code labels
autogate correct placement, B-1
adjusting movement, 6-32 label quality, B-3
extent, 6-33 label size, B-3
movement, 6-32 optical characteristics, B-6
autoloader orientation in carousel, 1-23
adding barcode labels, 1-22 print quality, B-3
barcode labels, 1-22 specifications, B-2, B-5
basic operating techniques, 1-21 symbol dimensions, B-3
carousel, 1-21 thickness, B-3
carousel install, 1-24 types read by the MCL, B-4
carousel remove, 1-26 used on sample carousel, 3-1
indicator light, 1-21 barcode labels
plate adapter install, C-1 autoloader, 1-22
plate adapter remove, C-4 bar-code reader
autoloader carousel acceptable bar codes, B-4
adding tubes, 1-23 MCL, specifications, B-6
automated software feature, 3-6 basic (with autoloader), 1-21
automatic export basic operating techniques
reports, 6-71 autoloader, 1-21
autosampling system batch analysis
Deep Clean solution peristaltic pump, 1-10 samples and tubes, 8-24
to cancel, 8-26
batch export
B reports, 6-69
backflush settings Batch export report
to change, 12-48
Index-2
Index
Hide Personal and Sensitive CDRH

Information, 6-71 define, Abbreviations-1
batch reports cell illumination
printing, 6-77 description, 3-4
batch review, 8-30 CFSE
cancel, 8-33 define, Abbreviations-1
BCI change
define, Abbreviations-1 password, 2-22
beam shaping changing
laser, 3-3 mixer and backflush settings, 12-48
BMP tube name, 6-16
define, Abbreviations-1 channel
bottle to set, 6-16
Deep Clean solution, 1-10 channels
BP adding for compensation, 7-18
define, Abbreviations-1 fluorescence, 6-45, 6-46
to set, 6-16
characteristics
C performance, 1-31
calculating check
compensation values, 7-9 waste and reagent levels, 4-2
concentration, 8-7 checksum algorithm
sample injection volume, 8-7 Codabar, B-7
calculation of the automatic compensation Code 39, B-7
experiment is incorrect, 10-18 Interleaved 2-of-5, B-7
calculations cleaning
compensation, 7-1 nonscheduled, 11-14
calibrate routine, 11-1
carrier (DxFLEX with autoloader), 12-50 sample probe, 11-8
calibrating sheath fluid container, 11-11
sample flow rate, 12-35, C-44 waste container, 11-12
cancel cleaning (without autoloader)
batch analysis, 8-26 sample station (without autoloader), 11-7
tube or sample review, 8-30 cleaning and ventilation
canceling installation environment validation, A-2
analysis time, 8-23 cleaning solution
batch review, 8-33 to prepare, 9-1
carousel cleaning the sample station (without
autoloader, 1-21 autoloader)
installing in autoloader, 1-24 routine maintenance, 11-7
removing from autoloader, 1-26 cleaning the sheath fluid container
carousel, sample routine maintenance, 11-11
bar-code labels, 3-1 cleaning the waste container
description, 3-1 routine maintenance, 11-12
See also MCL cm
carrier (DxFLEX with autoloader) define, Abbreviations-1
calibrate, 12-50 Codabar
carrier settings, 2-45 bar-code, B-5
caution label Codabar bar code
RoHS, 10-11 checksum algorithm, B-7
Index-3
Index
Code 128 main, 1-2

bar-code, B-5 optical, 1-3
Code 39 concentration
bar-code, B-5 calculation, 8-7
Code 39 bar code sample injection volume, 8-7
checksum algorithm, B-7 concentration calculation is incorrect, 10-19
collecting concluding
data, 6-47 experiment, 6-78
QC data, 5-8 configuration (with autoloader)
collection system, 1-19
acquisition screen, 2-5 configuration (without autoloader)
collection conditions system, 1-17
to set, 6-57 configuring
compensation acquisition settings, 6-53
adding channels, 7-18 confirming results, 5-12
calculations, 7-1 connection indicator light in the lower left
to adjust, 7-12 corner of the software screen is red and
to manually adjust, 7-12 displays Disconnected and Error, 10-12
compensation controls conventions used, 1-xxvii
compensation experiment screen, 2-11 copying
compensation experiment experiments, 8-1
to create, 7-3 previously acquired experiment, 8-1
compensation experiment screen, 2-11 report, 8-9
compensation controls, 2-11 create a compensation experiment
compensation library start page operations, 2-2
to manage, 7-17 create a new experiment from a template
compensation matrix start page operations, 2-2
to create from previously acquired create a new screen
data, 7-11 start page operations, 2-2
to generate, 7-6 creating
compensation sample compensation experiment, 7-3
prepare, 7-5 compensation matrix from previously
compensation settings acquired data, 7-11
adjust, 8-8 detector configuration, 6-44
exporting, 7-15 dot plot overlays, 8-5
importing and exporting, 6-74 heat map (plate loader), C-33
compensation settings from compensation Levy-Jennings charts, 5-15
library multi-data histograms, 8-5
to import, 7-14 new experiment, 6-1
compensation settings from compensation new panel experiment, 6-39
matrix files plots and gates, 6-19
to import, 7-13 report, 8-9
compensation values user roles, 2-24
to calculate, 7-9 users, 2-20
components creating auto gates, 6-30
Cytometer, 1-2 CSV
Fluid Containers, 1-2 define, Abbreviations-1
fluidics system, 1-8 custom statistics
instrument, 1-1 setting, 6-36
Index-4
Index
customer-replaceable parts data populations are normal on one laser, but

ordering, E-1 too low on another laser, 10-16
customized parameters data populations are not where they are
setting, 6-34 expected, 10-17
CV data sheets, material safety
define, Abbreviations-1 how to order, 1-31
cyber security, D-52 data storage
backing up your recovery key, D-13 description, 3-6
BitLocker decryption, D-17 database
BitLocker encryption, D-6 backup, 10-23
BitLocker keys, D-2 restore, 10-23
changing encryption strength, D-2 Deep Clean
disabling McAfee application control, D-48 procedure, 11-9
drive encryption, D-1 routine maintenance, 11-9
enabling McAfee application control, D-30 Deep Clean solution
exporting certificate, D-34 to add, 12-14
good practices in cyber security, D-1 to prepare, 12-14
Importing certificate, D-44 Deep Clean solution bottle
installing McAfee Application Control, D-26 fluidics module, 1-10
malware detection software, D-25 Deep Clean solution peristaltic pump
operating system patching, D-56, D-60 autosampling system, 1-10
recovering your BitLocker key, D-23 default
suspending BitLocker drive detector configuration, 6-45
encryption, D-21 report, 8-9
switching McAfee Application control to default password, 2-19
update mode, D-46 define
CytExpert certificate, D-34 %, Abbreviations-1
Cytometer, 1-2, 1-3 quality control (QC), 5-1
cytometer symbols, 1-vii
information defining the negative population, 7-6
viewing, 10-27 delete
Cytometer cannot be turned on, 10-12 heat map parameter, C-35
cytometer menu, 2-16 tubes, 1-23
delete page
report, 8-19
D deleting
daily clean heat map (plate loader), C-41
routine cleaning, 11-1 new panel, 6-42
daily clean (with autoloader-carousel mode) report, 8-9
routine cleaning, 11-4 standardization items, 5-29
daily clean (with autoloader-plate mode) user roles, 2-25
routine cleaning, C-41 users, 2-21
daily clean (without autoloader) designing
routine cleaning, 11-2 report, 8-10
data detector (WDM)
analyzing, 6-60 optical components, 1-3
collecting, 6-47 detector configuration
exporting, 6-60 to create, 6-44
sampling, 6-47 to edit, 6-44
to import, 8-1 to select, 6-44
Index-5
Index
to verify, 6-44 ventilation and cleaning, A-2

detector configuration default, 6-45 worktable, A-2
detector unit exit the software
wavelength division multiplexor (WDM), 1-4 start page operations, 2-2
dimensions experiment
instrument, 1-28 concluding, 6-78
disabling lasers, 6-54 experiment (new)
disposal to create, 6-1
precaution, 10-11 experiment saving, 6-78
waste, A-4 experiment selection
DNA start page, 4-26
define, Abbreviations-1 experiment settings, 2-41
dot plot overlays experiments
to create, 8-5 to copy, 8-1
downloading security updates, D-56 export
drive decryption batch reports, 6-69
enabling BitLocker, D-17 reports automatically, 6-71
drive encryption, D-1 exporting
BitLocker keys, D-2 compensation settings, 6-74, 7-15
BitLocker recovery, D-23 data, 6-60
changing encryption strength level for FCS files, 6-66
BitLocker, D-2 heat map (plate loader), C-41
enabling BitLocker, D-6 instrument settings, 6-73, 6-74
dual-parameter plots, 2-39 new panel, 6-41
plots of multiple tubes as picture files, 6-69
results, 8-34
E statistics table of multiple tubes as a picture
ECD file, 6-69
define, Abbreviations-1 user logs, 2-29
editing Exporting Batch Reports
detector configuration, 6-44 Hide Personal and Sensitive
standardization item parameters, 5-28 Information, 6-71
EFUP extent
define, Abbreviations-1 autogate, 6-33
electronic devices
fuse, 1-18, 1-20
load button, 1-18, 1-21 F
power switch, 1-18, 1-20 FAPD
USB connector for autoloader, 1-20 define, Abbreviations-1
USB connector for workstation, 1-20 FCS
EMF define, Abbreviations-1
define, Abbreviations-1 FCS files
EMR exporting, 6-66
define, Abbreviations-1 files menu, 2-15
enabling lasers, 6-54 filling sheath fluid container, 4-2
environment validation filter
installation, A-1 optical, 1-4
environment validation (installation) sheath fluid, 1-10
power source, A-3 FITC
temperature and humidity, A-3 define, Abbreviations-1
Index-6
Index
FL front cover
See fluorescent light (FL) removal, 12-2
flow cell FS
hydrodynamic focusing, 3-2 See forward scatter
illustration, 3-3 FSC
to prime, 12-34 define, Abbreviations-1
flow cell waste out functions
fluidic connection, 4-3 software, 2-1
Fluid Container holder, 1-9 fuse
Fluid Containers, 1-2, 1-3 electronic devices, 1-18, 1-20
component, 1-2 to replace, 12-43
fluidics system, 1-9
fluidics system component, 1-8
fluid sensor holder cutout, 1-9 G
fluid status information displays red for Sheath gain
and/or Waste even though the sheath to adjust, 6-54
fluid container is full and the waste gate settings, 2-44
container is empty, 10-13 gates
fluidic connections to create, 6-19
flow cell waste out, 4-3 gates for dual-parameter plots
sheath fluid in, 1-11, 4-3 four-quadrant gates, 2-40
sheath fluid level sensor connector, 1-11, 4-3 lasso gates, 2-40
sheath return, 1-11, 4-3 polygon gates, 2-40
waste level sensor connector, 1-11, 4-3 rectangle gates, 2-40
waste out, 1-11, 4-3 gates for single-parameter plots
Fluidics module line-segment gates, 2-40
alarm, 1-10 vertical gates, 2-40
fluidics module GB
Deep Clean solution bottle, 1-10 define, Abbreviations-1
fluidics system, 1-10 generating
fluidics system component, 1-8 compensation matrix, 7-6
sheath fluid filters, 1-10 Ghz
fluidics system define, Abbreviations-1
components, 1-8 Gr. Wt.
Fluid Containers, 1-9 define, Abbreviations-1
fluidics module, 1-10 graphic and gating styles, 2-39
fluidics system components graphics
Fluid Containers, 1-8 printing, 6-75
fluidics module, 1-8
fluorescence channels, 6-45, 6-46 H
fluorescent light (FL)
H
cell illumination, 3-4
define, Abbreviations-1
collection, 3-5
hazard
when to use, example, 3-4
laser, 10-1
focusing, hydrodynamic
hazards
description, 3-2
laser beam, 10-2
forward scatter (FS)
radiation, 10-2
cell illumination, 3-4
hazards/precautions, 10-1
four-quadrant gates
heat map (plate loader)
for dual-parameter plots, 2-40
Index-7
Index
creating, C-33 liquid flow tubing, 12-33

deleting, C-41 pre-boot, 4-1
exporting, C-41 inspection when unpacking, A-4
modifying, C-40 install
refreshing, C-39 carousel in autoloader, 1-24
heat map parameter plate adapter in autoloader, C-1
deleting, C-35 installation
help menu, 2-18 environment validation, A-1
Hide Personal and Sensitive Information software, A-5
Batch export report, 6-71 installation category, 1-28
sample property, 6-9, C-26 installing
histogram software, A-6
types of display, 3-7 installing security updates, D-60
humidity and temperature installing the instrument and connecting the
installation environment validation, A-3 equipment
hydrodynamic focusing unpacking inspection, A-5
description, 3-2 instrument
Hz components, 1-1
define, Abbreviations-1 initializing, 4-27
inspecting materials, A-4
maintenance, 12-1
I parameters, 1-29
I2-of-5 shut off, 9-1
bar-code, B-5 transportation and storage, A-1
IEC unpacking, A-4
define, Abbreviations-1 instrument characteristics, 1-28
importing instrument components
compensation settings, 6-74 Cytometer, 1-2
compensation settings from compensation Fluid Containers, 1-2
library, 7-14 instrument configuration
compensation settings from compensation unpacking inspection, A-4
matrix files, 7-13 instrument dimensions, 1-28
data, 8-1 instrument operations cannot be performed in
instrument settings, 6-73 the Acquisition screen, 10-20
lot-specific target values, 5-4 instrument settings
new panel, 6-40 exporting, 6-74
previously acquired data, 8-1 importing, 6-73
standardization item, 5-29 importing and exporting, 6-73
independent worksheets instrument specifications, 1-28
to switch, 6-15 instrument transport or storage
indicator light to prepare, 11-14
autoloader, 1-21 intended use, 1-1
information Interleaved 2-of-5
ordering, 1-27 bar-code, B-5
information for cytometer Interleaved 2-of-5 bar code
viewing, 10-27 checksum algorithm, B-7
initializing the instrument, 4-27 items
inspecting materials adding for standardization, 5-27
unpacking, A-4 deleting for standardization, 5-29
inspection
Index-8
Index
editing for standardization, 5-28 lenses

importing for standardization, 5-29 beam shaping, 3-3
IVD cross-cylindrical, 3-3
define, Abbreviations-1 Levey-Jennings charts
to create, 5-15
light collection, separation and measurement
K description, 3-4
kg line segment gates
define, Abbreviations-1 for single-parameter plots, 2-40
KO liquid flow tubing
define, Abbreviations-1 inspection, 12-33
liquid flow tubing inspection
L routine maintenance, 12-33
LJ
L
load button
label
electronic devices, 1-18, 1-21
RoHS caution, 10-11
log
labels
user management operation, 2-29
bar-code, specifications, B-5
logs
disposal of electrical instrumentation,
exporting, 2-29
warning, 10-10
viewing, 2-29
RoHS caution, 10-11
lot-specific target values
to set, 6-16
to import, 5-4
See also bar-code labels
LWH
labels, bar-code
See bar-code labels
language settings, 2-47
laser M
bar-code reader, MCL, B-6 m
beam shaping, 3-3 define, Abbreviations-2
hazards, 10-1 main components, 1-2
settings, 6-53 maintenance
standardizing target values, 5-26 instrument, 12-1
laser beam See also routine maintenance; nonscheduled
hazards, 10-2 maintenance
laser delay maintenance entry
to set, 12-40 adding, 2-30
laser delay values are out of range, 10-15 deleting, 2-30
laser lines, 1-29 maintenance log
laser power is low, 10-14 export, 2-37
laser wavelength, 1-29 maintenance entry, 2-30
lasers view, 2-37
disable, 6-54 maintenance logs
enable, 6-54 adding maintenence tasks, 2-32
lasso gates adding/deleting maintenance tasks, 2-32
for dual-parameter plots, 2-40 deleting maintenance tasks, 2-35
launching the software, 2-1 maintenance reminder
LED to manage, 12-11
define, Abbreviations-2 maintenance task
Index-9
Index
maintenance log, 2-32 user roles, 2-25

malware detection software, D-25 users, 2-21
managing movement
compensation library, 7-17 adjusting for autogates, 6-32
maintenance reminder, 12-11 autogate, 6-32
panel library, 6-38 MSDS (material safety data sheets)
manual how to order, 1-31
adjust compensation, 7-12 multi-data histograms
manual injection mode to create, 8-5
running small sample volumes, 4-9 multiple tube plots
master page export as picture files, 6-69
report, 8-9 multiple tube statistics table
material safety data sheets (MSDS/SDS)
how to order, 1-31 export as picture files, 6-69
MB mute
define, Abbreviations-2 alarm, 1-10
McAfee application certificate, D-44 mute alerter icon, 1-10
McAfee application control mW
disabling, D-48 define, Abbreviations-2
enabling, D-30
update mode, D-46
MCL (Multi-Tube Carousel Loader) N
loading the sample, 3-1 NA
menu define, Abbreviations-2
acquisition and analysis screen, 2-14 NaClO
advanced, 2-17 define, Abbreviations-2
cytometer, 2-16 NaN3
files, 2-15 define, Abbreviations-2
help, 2-18 navigation
QC, 2-17 acquisition screen, 2-4
setting, 2-16 QC screen, 2-13
software, 2-14 negative population
menu tree to define using unstained samples, 7-6
software, 2-14 new experiment
MFI to create, 6-1
define, Abbreviations-2 nm
Mhz define, Abbreviations-2
define, Abbreviations-2 no changes occurred after manually adjusting
min compensation settings, 10-17
define, Abbreviations-2 no data acquisition, 10-16
mixer is not functioning, 10-20 nonscheduled cleaning, 11-14
mixer settings nonscheduled maintenance
to change, 12-48 adding the Deep Clean solution, 12-14
mL calibrating the sample flow rate, 12-35, C-44
define, Abbreviations-2 cleaning the sample probe, 11-8
mm preparing the instrument for storage or
define, Abbreviations-2 transport, 11-14
modifying replacing the fuse, 12-43
heat map (plate loader), C-40 replacing the optical filter, 12-41
nonscheduled replacement/adjustment, 12-35
Index-10
Index
Nt. Wt. saving, 6-39

define, Abbreviations-2 parameter (heat map)
NW7 delete, C-35
bar-code, B-5 parameters
description, 3-7
editing standardization items, 5-28
O instrument, 1-29
open a compensation experiment TIME, 3-7
start page operations, 2-2 password
open an experiment change, 2-22
start page operations, 2-2 default, 2-19
open software, 4-4 reset, 2-22
opening PB
analysis screen, 8-3 define, Abbreviations-2
operating system patching, D-52 PC5
operating techniques, 1-21 define, Abbreviations-2
optical PC5.5
components, 1-3 define, Abbreviations-2
optical bench PC7
optical component, 1-3 define, Abbreviations-2
optical characteristics PE
bar-code labels, B-6 define, Abbreviations-2
for bar-code labels, B-6 PEEK
optical components define, Abbreviations-2
detector (WDM), 1-3 PerCP
optical bench, 1-3 define, Abbreviations-2
optical fiber, 1-3 performance characteristics, 1-31
optical fiber, 1-7 performing
optical filter, 1-4 daily quality control, 5-1
to replace, 12-41 performing standardization, 5-29
ordering PI
customer-replaceable parts, E-1 define, Abbreviations-2
ordering information, 1-27 picture files
output to export, 6-69
report, 8-21 plate adapter
overlay dot plots, 2-39 installing in autoloader, C-1
overlay histograms, 2-39 removing from autoloader, C-4
plot area
P acquisition screen, 2-8
plot display conditions
page setup settings, 2-45
to set, 6-59
panel
plot settings, 2-43
saving to panel library, 6-39
plots
to delete, 6-42
dual-parameter, 2-39
to export, 6-41
overlay dot plots, 2-39
to import, 6-40
overlay histograms, 2-39
panel experiment
single-parameter, 2-39
to create, 6-39
to create, 6-19
panel library
to link from worksheets to reports, 8-19
managing, 6-38
to set, 8-3
Index-11
Index
types of display, 3-7 flow cell, 12-34

PN printing
define, Abbreviations-2 batch reports, 6-77
policies graphics, 6-75
account, 2-26 procedure
pollution degree, 1-28 Deep Clean, 11-9
polygon gates
population amplitude is decreasing and CV Q
values are increasing, 10-15 QC
populations are drifting, 10-14 applying standardization, 5-32
position define, Abbreviations-2
sample tube holder, 1-13 laser target values, 5-26
power source result manager, 5-19
installation environment validation, A-3 QC aborted due to low event rate, 10-20
power source inspection QC data
pre-boot, 4-4 collecting, 5-8
power switch QC experiment screen, 2-11, 2-13
electronic devices, 1-18, 1-20 QC failed, 10-21
pre-boot inspection, 4-1 QC menu, 2-17
check waste and reagent levels, 4-2 QC menu tree
power source inspection, 4-4 software, 2-14
workstation connections inspection, 4-4 QC report screen, 2-11
precaution QC sample tube
disposal, 10-11 preparation, 5-3
precautions QC screen
safety, 1-vi navigation, 2-13
precautions/hazards, 10-1 QC software menu tree, 2-14
preparation process QC well plate
QC sample tube preparation, 5-3 preparation, C-18
QC well plate preparation, C-19 quality control
preparing perform daily, 5-1
compensation sample, 7-5 quality control (QC)
Deep Clean solution, 12-14 define, 5-1
instrument for storage or transport, 11-14
QC sample tube, 5-3 R
QC well plate, C-18
radiation
the cleaning solution, 9-1
hazards, 10-2
preparing QC sample tube
RAM
preparation process, 5-3
required materials, 5-3
rCV
preparing QC well plate
preparation process, C-19
reagent levels
required materials, C-18
check, 4-2
previously acquired data
recovery key, D-13
to import, 8-1
rectangle gates
previously acquired experiment
to copy, 8-1
refreshing
priming
heat map (plate loader), C-39
Index-12
Index
reinstallation procedures restore

right-side cover, 12-6 database, 10-23
removal procedures result manager
front cover, 12-2 QC, 5-19
right-side cover, 12-4 results
top cover of autoloader, 12-7 confirming, 5-12
remove to export, 8-34
carousel in autoloader, 1-26 review
plate adapter in autoloader, C-4 batch, 8-30
renaming cancel, 8-30
report, 8-9 cancel batch review, 8-33
replacement samples, 8-27
nonscheduled, 12-35 tubes, 8-27
routine, 12-2 RH
replacing define, Abbreviations-2
fuse, 12-43 right-side cover
optical filter, 12-41 reinstallation, 12-6
sample peristaltic pump tubing for sample removal, 12-4
loading, 12-20 RoHS
sheath fluid filter, 12-16 caution label, 10-11
sheath fluid harness and/or waste define, Abbreviations-2
harness, 12-45 role management, 2-23
replacing the peristaltic pump for sample roles
loading creating, 2-24
routine maintenance, 12-20 deleting, 2-25
replacing the sheath fluid filter modifying, 2-25
routine maintenance, 12-16 routine cleaning, 11-1
report, 8-9 daily clean, 11-1
add page, 8-19 daily clean (with autoloader-carousel
delete page, 8-19 mode), 11-4
linking plots from worksheets, 8-19 daily clean (with autoloader-plate
master page, 8-9 mode), C-41
output, 8-21 daily clean (without autoloader), 11-2
to copy, 8-9 routine maintenance
to create, 8-9 cleaning the sample station (without
to delete, 8-9 autoloader), 11-7
to design, 8-10 cleaning the sheath fluid container, 11-11
to rename, 8-9 cleaning the waste container, 11-12
to set as default, 8-9 Deep Clean, 11-9
zoom in/out, 8-19 liquid flow tubing inspection, 12-33
reports replacing the sample peristaltic pump
automatic export, 6-71 tubing for sample loading, 12-20
batch export, 6-69 replacing the sheath fluid filter, 12-16
printing, 6-77 routine replacement/adjustment, 12-2
required materials RPTM
QC sample tube preparation, 5-3 define, Abbreviations-2
QC well plate preparation, C-18 running
software installation, A-5 single positive control samples, 7-7
reset system startup program, 4-17, 4-21, C-10
password, 2-22 running small sample volumes
Index-13
Index
manual injection mode, 4-9 sample station

sample probe, 1-12
sample tube holder, 1-12
S to clean (without autoloader), 11-7
S/N washer station and mixer, 1-12
define, Abbreviations-2 sample station (cleaning without autoloader)
safety routine maintenance, 11-7
precautions, 1-vi sample station (with autoloader), 1-13
sample sample station (without autoloader), 1-12
analyze, 8-22 sample tube holder
checks before running, 6-43 sample station, 1-12
flow, description, 3-1 sample tube holder cannot move up and down
illuminated in the flow cell, 3-4 automatically, 10-13
loading, automated, 3-1 sample tube holder position
sample acquisition position sample acquisition position, 1-13
sample tube holder position, 1-13 sample loading position, 1-13
sample carousel standby position, 1-13
bar-code labels, 3-1 samples
description, 3-1 analyze, 8-21
See also carousel, sample and MCL review, 8-27
sample flow sampling
description, 3-1 data, 6-47
sample flow rate sampling flow rate is too fast, 10-14
to calibrate, 12-35, C-44 save
sample flow rate is unstable, 10-13 panel to panel library, 6-39
sample injection mode saving
selecting, 4-8 experiment, 6-78
sample is flowing, but no signal appears in the screen
plot, 10-18 acquisition, 2-3
sample loading analysis, 2-10, 8-1, 8-3
backup, 4-9 compensation experiment, 2-11
sample loading position QC experiment, 2-11, 2-13
sample tube holder position, 1-13 QC report, 2-11
sample peristaltic pump tubing software, 2-1
to replace, 12-20 SDS (safety data sheets)
sample peristaltic pump tubing for sample how to order, 1-31
loading selecting
to replace, 12-20 detector configuration, 6-44
sample peristaltic pump tubing for sample proper sample injection mode, 4-8
loading (replacing) set as default
routine maintenance, 12-20 report, 8-9
sample probe, 1-12 setting
sample station, 1-12 channel, 6-16
to clean, 11-8 channels, 6-16
sample probe is too low, 10-19 collection conditions, 6-57
Sample Property custom statistics, 6-36
Hide Personal and Sensitive customized parameters, 6-34
Information, 6-9, C-26 labels, 6-16
sample review laser delay, 12-40
cancel, 8-30
Index-14
Index
plot display conditions, 6-59 open, 4-4

the plots and statistics, 8-3 to start, A-11
setting menu, 2-16 update and reinstall, A-6
settings software installation, A-5
acquisition, 6-53 required materials, A-5
auto export, 2-47 starting the software, A-11
carrier, 2-45 software installation fails, 10-20
experiment, 2-41 software menu, 2-14
gate, 2-44 acquisition and analysis screen menu, 2-14
language, 2-47 software menu tree, 2-14
laser, 6-53 software operations
page setup, 2-45 create a new experiment, 2-2
plot, 2-43 software screen, 2-1
software, 2-41 start page, 2-2
tube, 2-41 start page operations, 2-2
sheath fluid container, 1-9 software settings, 2-41
to clean, 11-11 software update
to fill, 4-2 reinstalling CytExpert software, A-6
sheath fluid container (cleaning) specifications
routine maintenance, 11-11 bar-code labels, B-2, B-5
sheath fluid filter characteristics, 1-28
to replace, 12-16 instrument, 1-28
sheath fluid filter (replacing) specimen
routine maintenance, 12-16 See sample
sheath fluid filters SS
fluidics module, 1-10 See side scatter
sheath fluid harness and sensor, 1-9 SSC
sheath fluid in define, Abbreviations-2
fluidic connection, 1-11, 4-3 Standardization, 5-21, 5-29
sheath fluid level sensor connector , 5-21
fluidic connection, 1-11, 4-3 standardization, 5-20
sheath return applying in QC, 5-32
fluidic connection, 1-11, 4-3 calibrating the gain, 5-29
shut off laser target values, 5-26
the instrument, 9-1 Standardization beads
side scatter (SS) creating target medians, 5-21
cell illumination, 3-4 standardization item
collection, 3-5 adding, 5-27
when to use, example, 3-4 deleting, 5-29
signals importing, 5-29
processing of, operation principles, 3-6 standardization item parameters
single positive control samples editing, 5-28
to run, 7-7 standby position
single-parameter plots, 2-39 sample tube holder position, 1-13
SNR start page
define, Abbreviations-2 main software screen, 2-2
software selecting experiments, 4-26
functions, 2-1 start page operations
launching, 2-1 create a compensation experiment, 2-2
main screen, 2-1 create a new experiment from a
Index-15
Index
template, 2-2 troubleshooting, 10-1

exit the software, 2-2 alarm does not sound when the waste
main software screen, 2-2 container is full or the sheath fluid
open a compensation experiment, 2-2 container is low and the software status
open an experiment, 2-2 display is red, 10-12
starting calculation of the automatic compensation
the software, A-11 experiment is incorrect, 10-18
station (with autoloader) concentration calculation is incorrect, 10-19
sample, 1-13 connection indicator light in the lower left
station (without autoloader) corner of the software screen is red and
sample, 1-12 displays Disconnected and Error, 10-12
statistics Cytometer cannot be turned on, 10-12
setting, 3-8 data populations are normal on one laser,
to set, 8-3 but too low on another laser, 10-16
status bar data populations are not where they are
acquisition screen, 2-9 expected, 10-17
storing the instrument, A-1 fluid status information displays red for
suspending BitLocker drive encryption, D-21 Sheath and/or Waste even though the
switch sheath fluid container is full and the
to independent worksheets, 6-15 waste container is empty, 10-13
symbols instrument operations cannot be performed
define, 1-vii in the Acquisition screen, 10-20
system configuration (with autoloader), 1-19 laser delay values are out of range, 10-15
system configuration (without laser power is low, 10-14
autoloader), 1-17 mixer is not functioning, 10-20
system startup program no changes occurred after manually
to run, 4-17, 4-21, C-10 adjusting compensation settings, 10-17
no data acquisition, 10-16
population amplitude is decreasing and CV
T values are increasing, 10-15
table populations are drifting, 10-14
troubleshooting, 10-12 QC aborted due to low event rate, 10-20
target medians QC failed, 10-21
Standardization beads, 5-21 sample flow rate is unstable, 10-13
target values sample is flowing, but no signal appears in
laser standardization, 5-26 the plot, 10-18
temperature and humidity sample probe is too low, 10-19
installation environment validation, A-3 sample tube holder cannot move up and
test tube down automatically, 10-13
illustration in sample carousel, 1-23 sampling flow rate is too fast, 10-14
put into the carousel, 1-23 software installation fails, 10-20
test tubes table, 10-12
acquisition screen, 2-6 wash station drips during backflush, 10-19
threshold Workstation cannot be turned on, 10-12
to adjust, 6-55 tube
TIME parameter analyze, 8-22
description, 3-7 settings, 2-41
top cover of autoloader tube name
removal, 12-7 to change, 6-16
transporting the instrument, A-1
Index-16
Index
tube review creating, 2-20

cancel, 8-30 deleting, 2-21
tubes modifying, 2-21
adding to carousel in autoloader, 1-23 using unstained samples, 7-6
analyze, 8-21
delete, 1-23
review, 8-27 V
turn off V
auto recalculate, 6-31 define, Abbreviations-2
turn on VA
auto recalculate, 6-31 define, Abbreviations-2
turning the power on, 4-4 VAC
ventilation and cleaning
U installation environment validation, A-2
unlock verifying
user account, 2-22 detector configuration, 6-44
unpacking inspection, A-4 vertex
check instrument configuration, A-4 add to auto polygon gate, 6-31
installing the instrument and connecting vertical gates
the equipment, A-5 for single-parameter plots, 2-40
unpacking the instrument, A-4 viewing
unstained samples cytometer information, 10-27
to define the negative population, 7-6 user logs, 2-29
USB
USB connector for autoloader W
electronic devices, 1-20 W
USB connector for workstation define, Abbreviations-2
electronic devices, 1-20 wash station and mixer
USD-1 sample station, 1-12
bar-code, B-5 wash station drips during backflush, 10-19
USD-1.25 waste container, 1-9
bar-code, B-5 to clean, 11-12
USD-3 waste container (cleaning)
bar-code, B-5 routine maintenance, 11-12
USD-4 waste disposal, A-4
bar-code, B-5 waste harness
USD-6 to replace, 12-45
bar-code, B-5 waste level sensor connector
user account fluidic connection, 1-11, 4-3
unlock, 2-22 waste levels
user management, 2-18 check, 4-2
user management operation waste liquid harness and sensor, 1-9
log, 2-29 waste out
user roles fluidic connection, 1-11, 4-3
creating, 2-24 wavelength
deleting, 2-25 laser, 1-29
modifying, 2-25 wavelength division multiplexer (WDM), 1-4
users WDM
Index-17
Index
sample injection mode
selecting, 4-14
selecting
proper sample injection mode, 4-14
Workstation, 1-2, 1-3
Workstation cannot be turned on, 10-12
workstation connections inspection
pre-boot, 4-4
worktable
installation environment validation, A-2
Z
zoom in
report, 8-19
zoom out
report, 8-19
Index-18
Beckman Coulter, Inc.
Customer End User License Agreement
This Product contains software that is owned by Beckman Coulter, Inc. or its suppliers and is protected by
United States and international copyright laws and international trade provisions. You must treat the
software contained in this Product like any other copyrighted material. This license and your right to use the
Product terminate automatically if you violate any part of this agreement.
This is a license agreement and not an agreement for sale. Beckman Coulter hereby licenses this Software to
you under the following terms and conditions:
You May:
1. Use this software in the computer supplied to you by Beckman Coulter;
2. Maintain one copy of this software for backup purposes (the backup copy shall be supplied by Beckman
Coulter);
3. After written notification to Beckman Coulter, transfer the entire Product to another person or entity,
provided you retain no copies of the Product software and the transferee agrees to the terms of this
license agreement.
You May Not:

1. Use, copy or transfer copies of this Software except as provided in this license agreement;
2. Alter, merge, modify or adapt this Software in any way including disassembling or decompiling;
3. Loan, rent, lease, or sublicense this Software or any copy.
Limited Warranty
Beckman Coulter warrants that the software will substantially conform to the published specifications for the
Product in which it is contained, provided that it is used on the computer hardware and in the operating
system environment for which it was designed. Should the media on which your software arrives prove
defective, Beckman Coulter will replace said media free of charge within 90 days of delivery of the Product.
This is your sole remedy for any breach of warranty for this software.
Except as specifically noted above, Beckman Coulter makes no warranty or representation, either expressed
or implied, with respect to this software or its documentation including quality, performance,
merchantability, or fitness for a particular purpose.
No Liability for Consequential Damages

In no event shall Beckman Coulter or its suppliers be liable for any damages whatsoever (including, without
limitation, damages for loss of profits, business interruption, loss of information, or other pecuniary loss)
arising out of the use of or inability to use the Beckman Coulter Product software. Because some states do not
allow the exclusion or limitation of liability for consequential damages, the above limitation might not apply
to you.
General
This agreement constitutes the entire agreement between you and Beckman Coulter and supersedes any
prior agreement concerning this Product software. It shall not be modified except by written agreement
dated subsequent to the date of this agreement signed by an authorized Beckman Coulter representative.
Beckman Coulter is not bound by any provision of any purchase order, receipt, acceptance, confirmation,
correspondence, or otherwise, unless Beckman Coulter specifically agrees to the provision in writing. This
agreement is governed by the laws of the State of California.
C44966AB Warranty-1
Beckman Coulter, Inc. Customer End User License Agreement
Warranty-2 C44966AB
Related Documents
Operating Instructions
PN C44966
• Introduction
• System Overview
• Using the CytExpert Software
• Operation Principles
• Daily Startup
• Instrument Quality Control
• Data Acquisition and Sample Analysis
• Compensation
• Data Review
• Daily Shutdown
• Troubleshooting
• Cleaning Procedures
• Replacement/Adjustment Procedures
• Instrument Installation
• Barcode Specifications for the DxFLEX [With
Autoloader]
• DxFLEX [With Autoloader-Plate Mode]
• Good Practices for Cyber Security
• Table of Hazardous Substances
• Abbreviations
© 2020 Beckman Coulter, Inc.

All Rights Reserved

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Instructions for Use

Beckman Coulter Biotechnology (Suzhou) Co., Ltd.

© 2020 Beckman Coulter, Inc.

All other trademarks are the property of their respective

• Worldwide, find us via our website at

May be covered by one or more pat.- see

Beckman Coulter Eurocenter S.A.

Alerts for Danger, Warning, and Caution

Revision History, iii

CHAPTER 1: System Overview, 1-1

Putting Sample Tubes in a Carousel, 1-23

CHAPTER 2: Using the CytExpert for DxFLEX Software, 2-1

Changing a User Password, 2-22

CHAPTER 3: Operation Principles, 3-1

CHAPTER 4: Daily Startup, 4-1

CHAPTER 5: Instrument Quality Control and Standardization, 5-1

CHAPTER 6: Data Acquisition and Sample Analysis, 6-1

Printing Batch Reports, 6-77

CHAPTER 7: Compensation, 7-1

CHAPTER 8: Data Review, 8-1

CHAPTER 9: Daily Shutdown, 9-1

CHAPTER 10: Troubleshooting, 10-1

CHAPTER 11: Cleaning Procedures, 11-1

CHAPTER 12: Replacement/Adjustment Procedures, 12-1

Nonscheduled Replacement/Adjustment, 12-35

APPENDIX A: Instrument Installation, A-1

APPENDIX B: Barcode Specifications for the DxFLEX [With Autoloader], B-1

APPENDIX C: DxFLEX [With Autoloader -Plate Mode], C-1

APPENDIX D: Good Practices for Cyber Security, D-1

Suspending BitLocker Drive Encryption, D-21

APPENDIX E: Table of Hazardous Substances, E-1

1.1 Main Components [Without Autoloader], 1-2

3.4 Time vs Fluorescence plot, 3-7

1.1 WDM Optical Filter Mount Color Codes, 1-7

This introduction contains the following information:

• How to Use Your Manual

How to Use Your Manual

About this Manual

The information in your Instructions for Use manual is organized as follows:

CHAPTER 1, System Overview

CHAPTER 2, Using the CytExpert for DxFLEX Software

CHAPTER 3, Operation Principles

CHAPTER 4, Daily Startup

CHAPTER 5, Instrument Quality Control and Standardization

CHAPTER 6, Data Acquisition and Sample Analysis

CHAPTER 8, Data Review

CHAPTER 9, Daily Shutdown

CHAPTER 10, Troubleshooting

CHAPTER 11, Cleaning Procedures

CHAPTER 12, Replacement/Adjustment Procedures

APPENDIX A, Instrument Installation

APPENDIX B, Barcode Specifications for the DxFLEX [With Autoloader]

APPENDIX C, DxFLEX [With Autoloader -Plate Mode]

APPENDIX D, Good Practices for Cyber Security

APPENDIX E, Table of Hazardous Substances

This document uses the following conventions:

This chapter contains information on:

Figure 1.1 Main Components [Without Autoloader]

Figure 1.2 Main Components [With Autoloader]

NOTE The Autoloader is an optional accessory that can be purchased separately.