bio-rad chemiDoc mp imaging system

ChemiDoc™ MP Imaging System
with Image Lab Software
Instrument Guide
Version 6.0
ChemiDoc™ MP Imaging System
with Image Lab Software
Instrument Guide
Version 6.0
Bio-Rad Technical Support Department
The Bio-Rad Technical Support department in the U.S. is open Monday through Friday,
5:00 AM to 5:00 PM, Pacific time. Go to www.consult.bio-rad.com for worldwide
technical support.
Phone: 1-800-424-6723, option 2
Web: www.consult.bio-rad.com
Email: Support@Bio-Rad.com (U.S./Canada only)
For technical assistance outside the U.S. and Canada, contact your local technical
support office.
Notice
No part of this publication may be reproduced or transmitted in any form or by any
means, electronic or mechanical, including photocopy, recording, or any information
storage or retrieval system, without permission in writing from
Bio-Rad.
Bio-Rad reserves the right to modify its products and services at any time. This user guide
is subject to change without notice. Although prepared to ensure accuracy, Bio-Rad
assumes no liability for errors or omissions, or for any damage resulting from the
application or use of this information.
Credits
1. Image Lab software is based in part on the work of the Qwt project
(http://qwt.sf.net).
2. Image Lab software is based in part on the work of the CImg project
(http://cimg.sourceforge.net/). See license for details at:
http://www.cecill.info/licences/Licence_CeCILL-C_V1-en.html
3. Image Lab software is based in part on the work of the Independent JPEG Group
(http://www.ijg.org/)
GelStar is a trademark of FMC Corporation. IRDye is a trademark of LI-COR Biosciences.
Alexa Fluor, Coomassie Fluor, Qdot, OliGreen, PicoGreen, Pro-Q, SYBR, and SYPRO are
trademarks of Life Technologies Corporation. Coomassie is a trademark of BASF
Aktiengesellschaft. Cy2 and Cy3 are trademarks of GE HealthCare. DyLight and Krypton
are trademarks of Thermo Fisher Scientific Inc. GelGreen and GelRed are trademarks of
Biotium, Inc. Slo-Blo is a trademark of Littelfuse, Inc. Mitsubishi is a trademark of
Mitsubishi Companies.
Bio-Rad Laboratories, Inc. is licensed by Life Technologies Corporation to sell SYPRO
products for research use only under U.S. Patent Number 5,616,502.
Criterion Stain Free gels are covered by U.S. Pat. No. 7,569,130.
Copyright © 2017 Bio-Rad Laboratories, Inc.
Table of Contents
Safety and Regulatory Compliance. . . . . . . . . . . . . . . . . . . . . . . . . . . .7

Important Safety Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Warranty . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
General Precautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Regulatory Notices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Alert Icons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Cautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Warnings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
Instrument Safety Warnings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
Notice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Power Safety Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Voltage Setting Information. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Fuses. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Chapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13

ChemiDoc MP Imaging System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
System Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
CCD Camera and Lenses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Universal Hood III . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Image Lab Software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Emission Filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Optional Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Printer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Conversion Screens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Optional Light Sources . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Instrument Guide | iii

Table of Contents
Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Technical Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
For More Information. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Chapter 2 Acquiring Images. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

Creating Protocols. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Using Signal Accumulation Mode (SAM). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Accessing Protocol Setup Screens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Choosing an Application . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
Configuring a Single-Channel Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
Multichannel Protocols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Configuring a Multichannel Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Configuring Analysis Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Configuring Output Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Positioning the Gel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Running a Protocol with SAM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Saving SAM Images . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
Setting Up a Custom Application . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
Editing a Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
Chapter 3 System Calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Recalibrating the Imager . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
Appendix A Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
UV Transilluminator Lamp and Starter Replacement . . . . . . . . . . . . . . . . . . . . . . 45
Fuse Replacement. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Appendix B Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Appendix C Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Calibrating Accessories. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Installing Optional Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
iv | ChemiDoc MP Imaging System with Image Lab Software

Table of Contents
Epi Light Modules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51

UV/White Light Conversion Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
XcitaBlue Conversion Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
Gel Alignment Template Kit. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Ordering Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
Appendix D Using Bio-Rad Stain-Free Technology. . . . . . . . . . . . . 63

Stain-Free Workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Electrophoresis with Stain-Free Gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Imaging Gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Imaging Blots. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
Instrument Guide | v
Table of Contents
vi | ChemiDoc MP Imaging System with Image Lab Software

Safety and Regulatory Compliance
Important Safety Information

Please read these instructions before attempting to operate the ChemiDoc™ MP
imaging system.
This instrument is suitable for research use only. It must be used, therefore, only by
specialized personnel who know the health risks associated with the reagents that
are normally used with this instrument.
Use of the ChemiDoc MP imaging system involves UV illumination. Proper

precautions must be taken to avoid eye and skin exposure to the UV radiation. This
instrument is meant for use only by trained personnel who know the health risks
associated with the UV radiation normally used with this instrument. The acrylic
shield provides some UV protection. However, it does not guarantee complete
protection, and it is designed to shield only the person working in front of the
imager.
WARNING! Use of the acrylic screen does not guarantee the user protection
from UV radiation. The use of protective eyeglasses, mask, and/or gloves is
strongly recommended.
Instrument Guide | 7
Warranty
The ChemiDoc MP imaging system is warranted against defects in materials and
workmanship for one year. If any defect occurs in the instrument during this
warranty period, Bio-Rad Laboratories, Inc. will repair or replace the defective parts
at its discretion without charge. The following defects, however, are specifically
excluded:
 Defects caused by improper operation
 Repair or modification done by anyone other than Bio-Rad Laboratories,

Inc. or the company’s authorized agent
 Use of spare parts supplied by anyone other than Bio-Rad Laboratories,

Inc.
 Damage caused by accident or misuse
 Damage caused by disaster
 Corrosion caused by improper solvents or samples
General Precautions
 Read the user guide carefully.
 The instrument must be used only for the intended purpose of gel
documentation in research laboratories.
 The instrument must be connected to a grounded power source line and

protected by a circuit breaker.
 Do not pour liquids directly on or inside the instrument.
 Switch off all lights on the instrument immediately after use.
 Clean the transilluminator sample area after use.
8 | ChemiDoc MP Imaging System with Image Lab Software

Regulatory Notices
Regulatory Notices
The ChemiDoc MPimaging system is designed and certified to meet EN 61010, the
internationally accepted electrical safety standard, EMC regulations, and TUV
requirements. Certified products are safe to use when operated in accordance with
this user guide. Do not modify or alter this instrument in any way. Modification or
alteration of this instrument will
 Void the manufacturer’s warranty
 Void the regulatory certifications
 Create a potential safety hazard
Caution: Bio-Rad Laboratories, Inc. is not responsible for any injury or damage
caused by use of this instrument for purposes other than those for which it is
intended or by modifications of the instrument not performed by Bio-Rad
Laboratories, Inc., or an authorized agent.
Alert Icons
Alert icons in this guide call attention to caution and warning paragraphs. Each icon
indicates the type of hazard addressed.
Table 1. How alert icons are used in this guide

Icon Explanation
General
Indicates a potential hazard requiring special attention. This icon is used
when the hazard or condition is of a general nature.
Electrical hazard
Indicates a potential hazard requiring special attention when you are working
with electricity or electrical equipment.
Extreme heat and flammable materials

with extreme heat and flammable materials.
Radiation hazard
with UV radiation.
Cautions
A caution in this guide alerts you to take or avoid a specific action that could result
in loss of data or damage to the instrument. A caution can also indicate that, if the
precaution against a potential hazard is not taken, minor or moderate injury might
occur.
Example
Caution: With the exception of cleaning or replacing light bulbs, refer all
servicing to qualified Bio-Rad personnel or their agents.

Instrument Safety Warnings
Warnings
A warning in this guide precedes an action that, if not followed correctly, could
cause serious injury or death to the operator, serious or total loss of data, or serious
damage to the instrument.
Example
WARNING! Use of the acrylic screen does not guarantee the user protection
from UV radiation. The use of protective eyeglasses, mask, and/or gloves is
strongly recommended.
Instrument Safety Warnings

Before you operate the instrument, carefully read the contents of Table 2.
Table 2. Safety cautions and warnings for the instrument

Icon Meaning
Caution: With the exception of cleaning or replacing light bulbs, refer all
servicing to qualified Bio-Rad personnel or their agents. If you
experience technical difficulties with the instrument, contact Bio-Rad to
schedule service. The instrument should not be modified or altered in
any way. Alteration voids the manufacturer’s warranty and might create
a potential safety hazard for the user.
Caution: If the case interlock is defeated, there is a possibility of UV-B
radiation hazard due to UV-B light exposure. Exercise caution when
servicing the instrument.
Caution: Disconnect the AC power cord before removing the
instrument cover.
Warning: This instrument must be connected to an appropriate AC voltage

outlet that is properly grounded.
Notice
The ChemiDoc MP imagerimager is intended for laboratory use only. This device is
meant for use by specialized personnel who know the health risks associated with
reagents normally used in electrophoresis. Bio-Rad Laboratories, Inc. is not
responsible for any injury or damage caused by use of this instrument for purposes
other than those for which it is intended, or for instrument modifications not
performed by Bio-Rad Laboratories, Inc. or an authorized agent.
Power Safety Information
Voltage Setting Information

The universal hood for the ChemiDoc MP imager has a power supply that
automatically chooses the correct voltage for your country or region.
Fuses
The universal hood for the ChemiDoc MP imager has two user-serviceable fuses, F1
and F2, which are located on the bottom rear panel and are a part of the power
entry module. See Fuse Replacement on page 47 for more information.

1 Introduction
The ChemiDoc™ MP imaging system contains a charge-coupled device (CCD)

camera to capture images in real time and enable you to accurately position your
sample and generate optimized image data.
The ChemiDoc MP uses a new generation lighttight enclosure (the universal hood
III), which contains built-in UV and white light illumination as well as available red,
green, and blue epi LED light sources. The imaging system features a dynamic flat
fielding technology for superior image uniformity and accurate quantitation.
The imaging system includes the ChemiDoc MP instrument connected to a

separate computer running Image Lab™ software.
Using Image Lab software you can create protocols that specify options for single,
repeatable workflows that do the following:
 Control the ChemiDoc MP during image capture
 Optimize acquisition applications
 Analyze results
 Produce reports and other output
For more information about Image Lab software, see the Image Lab Software User
Guide.
1 | Introduction
ChemiDoc MP Imaging System

The ChemiDoc MP imaging system is a high-resolution gel documentation system
that allows fast, easy quantitation of gels and blots. Position your sample inside the
instrument and follow the on-screen steps in Image Lab software to run a protocol
with a single click. You can customize applications in an existing protocol or create
a new protocol using the options included in Image Lab software.
The imaging system also offers sensitive chemiluminescent detection. The system
includes a supersensitive CCD camera that is deeply cooled for faint-sample
detection and for accurate quantitation of image data.
Features include
 Smart, application-based protocol setup using Image Lab software, which

assists by presenting appropriate filter and illumination sources for imaging
applications that require excellent sensitivity
 Exceptional sensitivity and a dynamic range greater than four orders of

magnitude
 Flexibility to image chemiluminescent, fluorescent, and colorimetric

samples with dynamic flat fielding specific to each application*
System Components
CCD Camera and Lenses

The camera in the ChemiDoc MP imager is placed on top of a lighttight enclosure
(the universal hood) for capturing images. The camera comes with a motorized
zoom lens (MZL) that allows remote adjustment of the lens control functions (zoom,
focus, and iris).
A patented* software algorithm controls the MZL, giving the user automatic image
focus once an initial calibration is performed during system installation. Refer to
Technical Specifications on page 18.
* U.S. patent 8913127

System Components
A +1 diopter lens is factory installed to allow the entire sample stage to be visible.
This lens should always remain on the MZL assembly.
Universal Hood III

The universal hood III captures fluorescent and chemiluminescent images without
using a photographic darkroom. The enclosure has built-in white light epi-
illumination and UV transillumination. For easy sample loading, the UV
transilluminator is located in the drawer of the universal hood and can be accessed
from the front of the enclosure. When not imaging, the lights in the darkroom
enclosure turn off automatically.
The universal hood III has touchpad buttons to perform various functions. owever,
Image Lab software controls all of these functions remotely, removing any
requirement for manual control of the lens and lights. Running a protocol overrides
touchpad input.
Image Lab Software

The imager ships with a full version of Image Lab software. In addition to controlling
the imager, image capture, and optimization, Image Lab software can be used to
annotate and document images, analyze molecular weights when imaging protein
and nucleic acid gels, and determine quantitation and purity of samples.
You can print all or a subset of your data in a report. Alternatively, you can export
the data to other software, such as Microsoft Office programs, for further analysis or
presentation options.
Emission Filters
The universal hood III can hold up to four different emission filters for fluorescent
applications. No filter is required to image chemiluminescent samples.
There are six different filter wheel positions: four for the emission filters, one for a
calibration disc, and one that must be open for chemiluminescent applications. A
standard filter, included in the installation kit, is used for colorimetric (white light)
applications.
1 | Introduction
Optional Accessories
Bio-Rad Laboratories, Inc. offers a selection of optional filters and illumination
sources. See Ordering Information on page 60 for a complete listing of accessory
filters, UV light sources, optional parts, and replacement parts.
Printer
For your convenience, Bio-Rad offers an optional USB printer for use with the
ChemiDoc MP system: the Mitsubishi thermal printer (catalog #1708089).
Conversion Screens
White Light Conversion Screen
The white light conversion screen is a phosphor screen that produces white light
transillumination when placed on top of the UV transilluminator.
XcitaBlue Conversion Screen

The optional XcitaBlue™ screen kit (catalog # 1708182) converts UV to blue light,
which enables you to visualize a DNA sample while protecting it from UV damage.
Optional Light Sources

Red LED Module
The optional red LED module kit contains the emission filter and excitation source
for fluorescent applications. Instructions are also included.
Green LED Module

The optional green LED module kit contains the emission filter and excitation source
Blue LED Module

The optional blue LED module kit contains the emission filter and excitation source

Applications
The ChemiDoc MP imager is capable of running protocols to image blots that use
various detection reagents for chemiluminescent, colorimetric, and fluorescent
applications. It can also image singleplex, multiplex, and stain-free gels and blots.
Contact Bio-Rad technical support to determine whether your gel or blot can be
imaged on this instrument.
1 | Introduction
Technical Specifications
Applications
Chemiluminescence Yes
Fluorescence* Yes
Colorimetry Yes
Gel documentation Yes
Hardware Specifications
Maximum sample size  Length: 28 cm
 Width: 36 cm
Maximum image area  Length: 26 cm
 Width: 35 cm
Maximum image area for  Length: 25 cm
standard, UV-excited gels  Width: 26 cm
Excitation source  Trans-UV and epi-white are standard (302 nm included,
with 365 nm available as an option).
 Optional trans-white conversion screen.
 Optional XcitaBlue™ UV/blue conversion screen. Blue,
green, and red epis.
Detector Supercooled CCD
Pixel size (H x V in microns) 6.45 x 6.45
Cooling system Peltier cooled
Camera cooling –30°C controlled
temperature
Filter selector  6-position filter wheel
 1 without filter for chemiluminescence
Emission filters  1 included (standard)
 3 optional (530, 605, 695)
Dynamic range >4.0 orders of magnitude
Pixel density (gray levels) 65,535

Workflow
Dynamic flat fielding Application-specific, for all applications

Instrument size  Length: 36 cm
 Width: 60 cm
 Height: 96 cm
Instrument weight 32 kg
Operating Ranges
Operating voltage AC 110/115/230 V nominal
Operating temperature 10–28°C (21°C recommended)
Operating humidity <70% noncondensing
Automation Capabilities
Workflow automated Application driven, user-selected or recalled by a protocol
selection
Workflow automated Controlled by a protocol via application-specific setup for
execution image area, illumination source, filter, analysis, focus, and
reporting
Workflow reproducibility 100% repeatability via recallable protocols; from image
capture to quantitative analysis and reports
Autofocus Precalibrated focus for any zoom setting
Image flat fielding Dynamic; precalibrated and optimized per application
Autoexposure 2 user-defined modes (intense or faint bands
* Using the optional XcitaBlue kit (catalog # 1708182) is highly recommended if performing
preparative DNA applications with blue excitable stains. The UV to blue conversion screen allows
you to visualize DNA samples while protecting against UV damage.
Workflow
Following are the basic steps for acquiring, analyzing, and archiving an image using
the ChemiDoc MP imaging system and Image Lab software:
1. Select a protocol or customize a new one.
2. Position the gel or blot to be imaged.
3. Run your selected protocol.
1 | Introduction
4. View the displayed results.
5. Optimize the analysis.
6. Generate a report.
7. Save or export the results.
For More Information

For instructions about assembling and calibrating the ChemiDoc MP imager, refer to
the ChemiDoc MP Installation Guide in your ChemiDoc MP installation kit.
For information about installing Image Lab software, see Setting up Image Lab
Software in the Image Lab Software User Guide.
To recalibrate your imager when you acquire new accessories, see Chapter 3,
System Calibration.

2 Acquiring Images
Image Lab software runs configurable application-based, single-channel and

multichannel protocols for sample imaging. In a single acquisition, a protocol runs a
combination of settings for acquiring one or more images, analyzing them, and
creating a customized report.
Protocols can be retrieved, revised, and reused.
Images you acquire at high resolution result in large files. For this reason, Bio-Rad
recommends archiving images by exporting the files to network file server or to
removable storage media.
For more detailed information about protocols, see the Image Lab Software User
Guide.
2 | Acquiring Images
Creating Protocols
Creating a protocol consists of configuring acquisition, analysis, and output
settings. Analysis and output settings are optional. You can create a single-channel
or multichannel protocol.
To create a protocol, you configure these settings on the Protocol Setup screens.
The title bar above the Protocol Setup screen displays the protocol name and
imager type. Main steps appear as headings in the left pane of each screen.
Numbered steps appear under these headings.
Analysis settings consist of the following:
 Detecting lanes and bands
 Analyzing molecular weight
Output settings consist of the following:
 Printing the image
 Printing the report
 Displaying the report

Creating Protocols
To choose options for a protocol step, select the checkbox to the left of the
numbered step. To disable a numbered step, clear its checkbox.
Options for the selected step appear on the right side of the screen.
To determine the optimum imaging time for a chemiluminescent sample, select the
signal accumulation mode (SAM) option. SAM acquires a number of images with
different imaging times so you can compare the images and then select the
optimum imaging time.
Multichannel protocols acquire up to three separate images of a gel or blot, using

different illumination, filter, and exposure settings to create a combined image. The
Protocol Setup screen displays settings all three channels.
You can also use SAM in a multichannel protocol.
Using Signal Accumulation Mode (SAM)

When you run a chemiluminescence application, you can choose signal
accumulation mode (SAM) under Image Exposure on the Gel Imaging protocol
setup screen.
SAM simplifies obtaining an optimal image from a chemiluminescent sample. This

sample type often requires long integration times to obtain an image that represents
the best range of signal.
SAM presents a series of cumulative images with progressively greater signal in

each image. To run SAM you must estimate the shortest and longest times
expected to generate an image with the appropriate signal intensity. You then
decide how many total images to acquire in this period of time.
For example, if the minimum expected time to image the sample is 1 minute and the
maximum is 5 minutes, you enter these values (in seconds) in the Setup dialog box.
The value you enter in the Total number of images box defines the number of
images SAM captures in the defined interval.
In this example, images will be acquired at 1 minute intervals, starting at 1 minute

and ending at 5 minutes. The second 1-minute image is added to the first 1-minute
image, and the final image is the result of integrating these two images. The third 1-
minute image is added to the previous image, and so on, until the last image is
presented.
Although SAM is useful for determining the optimum imaging time for a
chemiluminescent sample, it results in data that are not as accurate as data from a
single image. Signal that is near the intensity of background noise becomes
increasingly masked as the number of cumulative images grows. To identify
extremely faint signals in an image, reacquire it as a single image, using the time the
SAM tool found to be appropriate.
Note: For more information about acquiring and saving SAM images, see
Running a Protocol with SAM in this chapter.

Creating Protocols
Accessing Protocol Setup Screens

To access the default Protocol Setup screen
1. Verify that the name of your imager appears on the status bar.
The green check mark indicates that the imager and the computer running
Image Lab software are communicating with each other.
2. Click New Protocol on the toolbar.
3. Select New Single Channel Protocol in the dropdown menu.
The Protocol Setup screen appears. Gel Imaging is selected in the left pane.
Gel imaging options appear on the right side of the Protocol Setup screen. The
protocol name appears in the title bar. You can change this name when you
save the protocol.
Choosing an Application
The term application refers to sample type. The sample types consist of the
following options:
 Nucleic acid gel, protein gel, or blot
 Detection reagent — dye or stain
In the Gel Imaging Protocol Setup screen, you can choose from a list of predefined
applications for common sample types. Each sample type is predefined with
optimal acquisition settings.
To use a dye or stain not listed in the Application dropdown menus, you can choose
the application type Custom and create a custom application. For more information,
see Setting Up a Custom Application on page 38.
Configuring a Single-Channel Protocol

 On the Protocol Setup screen toolbar, click New Protocol and select New
Single Channel Protocol.
The Protocol Setup screen appears with Gel Imaging selected. Acquisition
options appear in the right pane.
To configure the acquisition settings

1. Under Application, click Select an application type.
2. In the dropdown menu that appears, choose a predefined application.

Configuring a Single-Channel Protocol
Note: When you select Stain Free Gel, you can also select the gel
activation time. See Appendix D, Using Bio-Rad Stain-Free Technology, for
more information.
 Under Imaging Area, select the appropriate gel type or enter image area
dimensions. The red box to the right represents the imaging area for the
selected gel. The gray rectangle represents the imager sample stage.
Under Image Exposure, select one of the following:
 Auto Exposure — estimates an optimal exposure time and ensures the

best use of the dynamic range.
 Intense Bands — optimizes exposure for all bands.
 Faint Bands — uses a longer exposure time, making faint bands more
visible; more prominent bands might be overexposed.
After imaging a gel optimized for automatic exposure, the exposure time
used appears on screen. You can use it as a reference to set a manual
exposure time.
 Manual Exposure — overrides the automatic option. Exposure time can

range from 0.001–7,200 seconds. You can view the image exposure time in
the Image Info dialog box, which you can access in the Display Toolbox
above the on-screen image.
3. (Optional) Signal Accumulation Mode — when you run a chemiluminescence

application, you can also select SAM. To do so, select it under Image Exposure.
4. Display Options — two settings enable you to fine-tune image appearance:
 Highlight saturated pixels — display saturated pixels in red, which

indicates how much of the gel image is saturated. You can change this
option later: select View > Image Transform.
 Image color — select a color scheme for the sample image display.
Viewing the image with a different color scheme can make all of its
elements more visible. See the chapter, Viewing Images, in the Image Lab
Software User Guide for more information.
5. Save the protocol and run these options or go to the analysis or output
settings.

Multichannel Protocols
You can create a multichannel protocol, which sequentially acquires up to three
separate images of a gel or blot, using different illumination, filter, and exposure
settings for each image and also displays a composite of the images. You must
configure acquisition settings for at least two channels to create a valid
multichannel protocol.
The first Protocol Setup screen, Gel Imaging, presents different predefined
application options and acquisition settings for up to three channels.
Configuring a Multichannel Protocol

To access the Protocol Setup screen
1. Verify that the imager name appears on the status bar with a green check mark.
This indicates that the imager is communicating with Image Lab.
2. Click New Protocol on the toolbar and select New Multichannel Protocol.
The Protocol Setup screen for a multichannel image appears.
In the left pane, step 1, Gel Imaging, is already selected.
To configure acquisition settings

1. In the right pane of the Protocol Setup screen, click Configure in the Channel 1
box.
The Configure Channel dialog box appears.
2. Under Application, click Select and choose a predefined application in the

menu that appears.

Note: When you select Stain Free, you can also select the gel activation
time. See Appendix D, Using Bio-Rad Stain-Free Technology, for more
information.
3. Under Image Exposure, select one of the following:
 Auto Exposure — estimates an optimal exposure time and ensures the

best use of the dynamic range.
 Intense Bands — optimizes exposure for all bands.
 Faint Bands — uses a longer exposure time, making faint bands more
visible; more prominent bands might be overexposed.
After imaging a gel optimized for automatic exposure, the optimized

exposure time appears. You can use it as a reference to set a manual
exposure time.
 Manual Exposure — manually set an exposure time from 0.001–7,200

seconds.
Note: You can view the image exposure time in the Image Info dialog
box, which you can access in the Display Toolbox above the acquired
image.
 Signal Accumulation Mode — when you run a chemiluminescence

application, you can also use signal accumulation mode (SAM).
To do so, click Signal Accumulation Mode and select Setup to display the
Signal Accumulation Setup dialog box.
4. Set the following attributes under Display Options:
 Highlight saturated pixels — display saturated pixels in red, which

indicates how much of the gel image is saturated. You can change this
option later by selecting View > Image Transform.

 Channel Color — select a color to display the sample image. Assigning

each channel a different color makes it easy to identify the channels.
After you set up the first channel, the second channel box becomes active.
5. Repeat steps 2–4 to set up the remaining channels. The software lists
applications that remain available after each selection.
6. (Optional) To change channel settings, click Configure in the channel box and
select new settings.
7. Under Imaging Area, select a Bio-Rad gel or enter image dimensions. The red
line in the box depicts the imaging area for the gel. The gray rectangle depicts
the sample stage.
8. Save the protocol and run it or go to the analysis or output settings.
Configuring Analysis Settings

To analyze a gel or blot automatically, configure analysis settings in the following
protocol setup tasks:
 Detect Lanes and Bands
 Analyze Molecular Weight
For information about these settings, see the chapter, Protocols, in the Image Lab
Software User Guide.
Configuring Output Settings

When you display a report, a scrollable report screen appears in which you can view
the image, acquisition settings, and analysis data. You can view or print a single
image or report.
Note: You cannot print a report on a thermal printer.
To specify protocol output

1. Select Specify Output in the left pane of the Protocol Setup screen.
Output options appear in the right pane.

Positioning the Gel
2. Choose one of the following:
 Automatically print the image
 Automatically print a report
 Display the report
Image Lab prints to the default printer unless you select another printer.
See the chapter, Generating Reports, in the Image Lab Software User Guide for
information about customizing reports.
Positioning the Gel

After configuring protocol setup steps, position the gel on the imaging stage and
adjust the camera zoom level in Image Lab.
With the Bio-Rad gel alignment template kit you can center four sizes of standard
agarose gels and ensure the consistent placement of each gel. See Appendix C,
Accessories for more information.
Tip: To review protocol settings, click Protocol Summary in the left pane.
To position a gel
1. Place a gel on the imaging stage and center it.
2. In Image Lab, click Position Gel in the Protocol Setup screen.
3. The Run screen appears.
4. Adjust the camera zoom level with the slider below the image.
5. Click Run Protocol.
After the protocol runs, the acquired image appears.

Running a Protocol with SAM
Running a Protocol with SAM

Note: SAM can be used only with chemi applications.
You can interrupt the acquisition of images for a SAM-enabled protocol at any time.
To do so, click Stop Acquire and Continue with Selected. The acquisition process
stops and then continues the protocol with the image you select. Images already
acquired are discarded.
To view a SAM image in the workspace

 Click its thumbnail.
To save the image you want to analyze

1. Review the images and identify the image you want to use in your analysis.
Important: Save the image you want to keep before you continue running
the protocol. When you click Select Image and Continue, the selected
image is saved and all unsaved images are deleted.
2. Select the image you want and click Select Image and Continue.
Image Lab continues to the next step in the protocol using the image you
selected.
Saving SAM Images

To save a SAM image
1. Right-click its thumbnail and click Save.
2. In the Save File dialog box, accept the default file name or enter another name.
Click Save.
To save all SAM images

1. Right-click a SAM image and select Save All.
2. In the Select Directory dialog box, enter a folder name and click Choose.
The images are saved in the specified folder. The name of the file includes the
user name, timestamp, and exposure time. For example:
John Doe 2015-05-01 15 hr 44 min_Exposure_5.0sec.
Setting Up a Custom Application

When you create a protocol, Image Lab displays a list of application types and
predefined applications for acquiring images. To use a dye or stain not listed in the
Application dropdown menus, you can choose the application type Custom and
create a custom application. Also choose Custom to do the following:
 Choose a previously saved custom application
 Edit or rename a custom application
Note: If you are not sure how to configure a custom application for a dye or
stain, contact Bio-Rad Technical Support.
To create a custom application

1. On the Protocol Setup screen, select 1. Gel Imaging.
2. Under Application, click Select and choose Custom on the menu that appears.

Setting Up a Custom Application
The Manage Custom Applications dialog box appears.
3. Click New.
4. Enter a unique application name.
5. Under Application Settings, select a light source, filter, and image color.
Viewing the image with a different color scheme can make all of its elements
more visible.
6. Select a binning setting.
A higher binning setting combines pixels to increase the amount of signal

without increasing noise. A higher setting provides optimal sensitivity for low-
light applications such as chemiluminescence, but it reduces image resolution.
Editing a Protocol
You can open a protocol, change its settings, and save it with another name. You
can also disable a step in the saved protocol. When you edit a default protocol and
save it with a new name, the default protocol is unaffected.
See the chapter, Protocols, in the Image Lab Software User Guide for more
information.

3 System Calibration
When the imager is installed, it is calibrated using a calibration wizard. For detailed
instructions, see the installation guide in the imager installation kit.
The Image Lab™ software instrument calibration wizard provides several options
required to automate the system and prevent focus problems. Each of these
calibrations affects the system as follows:
 Focus Calibration — allows automated focus settings at any zoom point,

using a software algorithm. This ensures that the focus remains correct
whether you view an entire sample or an area of interest.
 Focus Calibration with Height Offset —takes the tallest of the available
conversion screens into account and extrapolates values for the others so
that focus remains optimal for the screen in use. Checking the Illumination
options on the Instrument Setup screen instructs Image Lab to generate
the focus calibration offset. Focus calibration offset does not change flat
field calibrations.
 UV Flat Field Calibration —generates the flat field correction profiles required
for the UV light source. The orange flat field generates a flat field profile for UV.
Because of this calibration, the images have more accurate quantity reporting
and backgrounds of even intensity.
 Lens Flat Field Calibration — corrects for the intensity roll-off inherent in
any lens.
 White Conversion Screen Calibration —generates a flat field correction

profile required for the white light conversion screen.
3 | System Calibration
Recalibrating the Imager

When you add light sources or filters to the ChemiDoc™ MP imager, you must
recalibrate the imager.
To recalibrate the imager

1. In Image Lab, click Edit > Instrument Setup.

Recalibrating the Imager
2. The Instrument Setup dialog box appears.
3 | System Calibration
3. (Optional) To add a new light conversion screen, select the appropriate box in
the Illumination Options field.
For more information, see Appendix C, Accessories.
4. For any other changes to the optical pathway, perform a flat field calibration. To
do so, click Reset in the Instrument Calibration > Flat Field group and follow the
on-screen instructions.
5. Wait for the software to prompt you to recalibrate the new illumination sources.
6. Click OK.

A Maintenance
UV Transilluminator Lamp and Starter Replacement

Note: Always keep the UV filter surface clean from the chemical agents used
as gel dyes. Use protective gloves when touching the UV transilluminator cover.
Depending on usage, the UV bulbs and starters can last for many years. Replace
bulbs when you notice them flickering. If a bulb does not turn on when it is new or
moved, replace the bulb starter and test the bulb again.
Three types of bulbs are available. The catalog numbers are listed in Ordering
Information on page 60. The standard bulb is 302 nm. Optionally, the 254 nm bulb is
used for cross-linking protein, and the 365 nm bulb is used to minimize denaturing
of DNA.
To replace the lamps

1. Turn off the power.
2. Disconnect the power cord from the universal hood.
3. Remove the four screws located on the left/right sides of the transilluminator
cover.
4. Remove the cover with the UV glass by sliding it forward, then lifting up.
5. Place it on a nonabrasive surface so that the glass does not get scratched or
damaged.
Note: Do not put the UV cover directly on the bench. Wear gloves when
touching the lamps.
A | Maintenance
6. Rotate the lamp until it becomes loose and the pins come to a vertical position.
7. Remove the lamp. Install the new lamp by rotating so that the pins are
horizontal and the lamp is tight.
8. Remove the starter by rotating it counterclockwise, and then pull it out.
9. Insert a new starter into the holder and rotate clockwise.
10. .Reassemble the cover and retighten the screws on both sides.

Fuse Replacement
Fuse Replacement
Always unplug the instrument before changing or checking the fuses.
Fuse holders
This unit is protected by two fuses (5 x 20 mm, 2 A Slo-Blo). The fuses are located in
fuse holders housed in the power entry module. This module is located on the right
side of the back of the universal hood.
To replace the fuses

1. Unplug the main power cable from the power outlet.
2. Use a flat screwdriver to turn the slotted front of each fuse holder
counterclockwise; the holder pops out so you can extract the fuse.
3. Remove the blown fuses and replace them with two new fuses
(catalog #9008935).
4. Slide each fuse holder into the power entry module until it snaps in place.
A | Maintenance

B Troubleshooting
The following table lists potential problems and suggested solutions.
Problem Possible Cause Solution

Camera does not  Power to the camera  Turn on the power to the camera.
respond/camera not may be turned off.
found  The camera cables  Make sure that all cables are
might not be seated connected as shown in the
properly. Installation Guide.
 If the camera driver is not present,
 The software driver for reload the camera driver from the
the camera is missing. Image Lab™ software CD.
 Disable the power-saving modes on
 Computer power-saving the computer.
modes might be
interfering with the
camera driver.  Replace the cables.
 The cables might be
defective.  Replace the camera.
 The camera might be
defective.
Horizontal stripes in  The emission filter might  Cycle power to the Universal Hood III
image when using not be positioned so that the filter wheel is positioned
the UV mode properly. properly.
Image is not visible  The monitor settings are  See your computer manual for the
on the monitor incorrect. proper settings.
 The lens cap is  Remove the lens cap.
attached.
B | Troubleshooting
Problem Possible Cause Solution

Printout does not  The monitor settings are  See your monitor manual for the
look like the monitor wrong. appropriate settings.
image
 The printer settings are  See your printer manual for the
wrong. appropriate settings.
Light leakage into the  The lens body is not  Loosen the thumbscrew and seat the
darkroom seated properly against lens properly against the gasket on
the gasket on the hood’s the hood’s adapter plate.
adapter plate.
Unable to focus on  Focus is not calibrated  Select Edit > Instrument Setup to
the sample using for samples using this recalibrate the focus for use with this
white light light source. accessory.
conversion screen
Lens limits seem  The camera lens is not  Re-seat the camera on the lens
artificially restricted seated properly on the mounting plate.
lens mounting plate.

C Accessories
Calibrating Accessories
When you install accessories during initial installation of the imaging system, you
must run the one-time Instrument Calibration wizard to calibrate the system. Refer
to the installation guide for instructions.
You must also recalibrate the imager before using new conversion screens, light
sources, or filters. See Chapter 3, System Calibration for more information about
calibrating new accessories.
Installing Optional Accessories
Epi Light Modules

Epi light modules are available for the ChemiDoc™ MP imager in three colors: red
(catalog #1708283), blue (catalog #1708285), and green (catalog #1708284). For
installation instructions, see Installing an Epi Light, an instruction sheet that
accompanies each module.
C | Accessories
UV/White Light Conversion Screen

This optional white light conversion screen (catalog #1708289) converts the UV light
generated in the universal hood to white light. The imager must be calibrated to use
the white light conversion screen.
The UV/White Light conversion screen is held in place by adhesive-backed edge

guides. After the edge guides are installed, the conversion screen remains centered
and does not slide, even when you close the drawer rapidly.

To install the UV/White Light conversion screen

Important: Do not remove the adhesive paper tape from the edge guides until
step 2. You cannot reposition the adhesive surfaces of the edge guides once
they are set in place.
1. Holding the screen right side up with the paper taped edge guides underneath,
practice positioning the edge guides so that they touch the inside of the
transilluminator drawer front and fit over the edge of the metal transilluminator
border (shown in red).
Adhesive-backed
edge guide
Transilluminator
drawer front
Transilluminator
border edge
2. When you have positioned the edge guides to your liking, remove the paper
tape from under each guide.
3. Carefully press each edge guide into position.
4. To visualize a sample using the screen, place the screen between the edge
guides.
5. Use the gel alignment template kit to center the gels on top of the screen
consistently. For more information, see To use a gel alignment template with a
conversion screen on page 58.
C | Accessories
To calibrate the imager

1. In Image Lab™ software, choose Edit > Instrument Setup.
2. Select the White Light Conversion Screen checkbox under Illumination

Options.
3. Under Instrument Calibration, click Reset for Focus Calibration.
When calibration finishes, the imager is ready to acquire images.
To image a gel with the white light conversion screen

1. Center the conversion screen on the imager stage.
2. Center the samples on top of the conversion screen.
3. Image the gel.
For more information, see Chapter 3, Acquiring Images.

XcitaBlue Conversion Screen

The optional XcitaBlue™ conversion screen kit (catalog #1708182) converts UV to
blue light, so you can visualize DNA samples while protecting them against UV
damage.
The XcitaBlue conversion screen is held in place by adhesive-backed edge guides.

After the edge guides are installed, the conversion screen remains centered and
does not slide even when you close the drawer rapidly.
Note: After you install the conversion screen, you must calibrate the imager.
To install the XcitaBlue conversion screen

Important: Do not remove the adhesive paper tape from the edge guides until
step 2. You cannot reposition the adhesive surfaces of the edge guides once
they are set in place.
1. Holding the screen right side up with the paper taped edge guides underneath,
practice positioning the edge guides so that they touch the inside of the
C | Accessories
transilluminator drawer front and fit over the edge of the metal transilluminator
border (shown in red).
Adhesive-backed
edge guide
Transilluminator
drawer front
Transilluminator
border edge
2. Position the edge guides to your liking.
3. Remove the paper tape from under each guide.
4. Carefully press each edge guide into position.
5. To visualize a sample using the screen, place the screen between the edge
guides.
6. Use the gel alignment template kit to center the gels on top of the screen
consistently.
To calibrate the imager

1. In Image Lab software, choose Edit > Instrument Setup.
2. Under Illumination Options, select the XcitaBlue Conversion Screen checkbox.
3. Under Instrument Calibration, click Reset for Focus Calibration.

When calibration finishes, the instrument is ready to acquire images.
To image a gel on the blue light conversion screen

1. Center the conversion screen on the imaging stage.
2. Center the samples on top of the conversion screen.
3. Select a protocol and image the gel.
For more information, see Chapter 3, Acquiring Images.
Gel Alignment Template Kit

The Bio-Rad gel alignment template kit (catalog #1708184) enables you to center
four sizes of standard agarose gels quickly and ensures the consistent placement of
each gel.
Magnetic
locator
frame
7 x 7 cm 15 x 7 cm 7 x 10 cm
15 x 10 cm
Note: Using the gel alignment template kit does not affect imager calibration.
The kit contains
 Magnetic locator frame
 Instruction sheet
 Alignment guides for the following gel trays:
C | Accessories
 Sub-Cell® GT UV-transparent mini-gel tray, 7 x 7 cm
 Sub-Cell GT UV-transparent wide mini-gel tray, 15 x 7 cm
 Sub-Cell GT UV-transparent mini-gel tray, 7 x 10 cm
 Sub-Cell GT UV-transparent gel tray, 15 x 10 cm
The gel alignment templates fit exactly into the XcitaBlue conversion screen frame
(catalog #1708182).
To use a gel alignment template with a conversion screen

1. Remove the magnetic locator frame if it is on the transilluminator.
2. Place the conversion screen on the transilluminator.
3. Place the gel alignment template that matches the size of the sample tray or
agarose gel inside the conversion screen frame.
4. Place the gel or gel tray into the open area of the template.
To use a gel alignment template with the magnetic locator frame

1. Place the magnetic locator frame over the transilluminator with the magnetic
side down.
2. Match the corners of the magnetic locator frame with the edges of the
transilluminator.
3. Position the magnetic locator frame so that its UV symbol matches the
direction of the UV symbol on the imager.
For example:
4. Place the gel alignment template that matches the size of the sample tray or
agarose gel into the magnetic locator frame.
5. Place the gel or gel tray into the open area of the template.

1.
The Instrument Setup dialog box appears.
2. Under Flat Field, click Reset and then click OK.
C | Accessories
Ordering Information
The following table lists catalog numbers and descriptions for all parts available for
the ChemiDoc MP imager, plus all optional accessories and replacement parts. For
more information, see the Bio-Rad catalog.
Table 3. Ordering information

Catalog # Description
1708280 ChemiDoc MP imaging system
Installation Kits
1708282 ChemiDoc MP installation kit
Universal Hood
1708281 Universal Hood III
Imaging Cameras
1708255 ChemiDoc MP camera with motorized zoom lens
Image Lab Software
1709690 Image Lab software, Windows/Mac
1708001 UV/White light conversion screen (UV to white light)
1708182 XcitaBlue (UV to blue light) conversion screen kit, without standard detection filter
1708283 Kit, Red LED Module
1708284 Kit, Green LED Module
1708285 Kit, Blue LED Module
1703759 Bio-Rad fluorescent ruler
1703760 Gel cutter ruler
1708184 Gel alignment template kit
Replacement Parts
1708026 Image Lab focus calibration target
1708027 Image Lab flat fielding disc
1708185 XcitaBlue viewing goggles
1707813 Sample holders for gels
1708081 Filter, standard emission, 62 mm
Lamps
1001361 UVB lamp, 302 nm (1 each)
1708097 302 nm lamp kit, (6 lamps)
1706887 365 nm lamp kit, (6 lamps)

Ordering Information
Table 3. Ordering information, continued

Catalog # Description
Fuses
9008935 Fuse T 2 A, 250 V, quantity 10
9000234 Fuse T 4 A, 250 V, quantity 10
Universal Hood III
1002787 Universal Hood feet, quantity 4
1708068 UV Shield for Universal Hood
Connection Cables
9310071 Cable, USB, Type A to B, 10 ft
9010064 Cable, USB, Type A to B, 6 ft
C | Accessories

D Using Bio-Rad Stain-Free
Technology
Bio-Rad stain-free gels eliminate the time-consuming staining and destaining steps
required by other protein detection methods. Stain-free gels include unique trihalo
compounds that allow rapid fluorescent detection of proteins with the ChemiDoc™
MP imager without staining.
When using Image Lab™ software, the ChemiDoc MP imager is stain-free enabled
to image the following gels:
 Criterion™ TGX Stain-Free™ precast gels
 Criterion Stain Free™ precast gels
 Mini-PROTEAN® TGX Stain-Free™ precast gels
 TGX Stain-Free™ FastCast™ acrylamide solutions for handcast gels
When trihalo compounds in the gels encounter tryptophan residues, a UV

light-induced reaction produces fluorescence, which can be easily detected by the
imager in gels or on low fluorescence polyvinyl difluoride (PVDF) membranes.
Activation of the trihalo compounds in the gels adds 58 Da moieties to available
tryptophan residues and is required for protein visualization. Proteins that do not
contain tryptophan residues cannot be detected using this technology. The
sensitivity of stain-free gels is comparable to staining with Coomassie Brilliant Blue
for proteins with a tryptophan content >1.5%; sensitivity superior to Coomassie
staining is possible for proteins with a tryptophan content >3%.
The benefits of stain-free technology include
 Elimination of staining and destaining steps for faster time to results
D | Using Bio-Rad Stain-Free Technology
 No background variability within a gel or between gels (as is often seen

with standard Coomassie staining)
 Elimination of the need for acetic acid and methanol in staining and
destaining, which reduces organic waste
 Visualization of transferred or blotted proteins on low fluorescence PVDF

membranes
Stain-Free Workflow
For detailed information about the Activate/image gels step, refer to Chapter 3,
Acquiring Images. For all other workflow steps, refer to the Criterion™ Precast Gels
Instruction Manual and Application Guide (bulletin #4110001) or to the
Mini-PROTEAN® Precast Gels Instruction Manual and Application Guide
(bulletin #1658100).
Prepare buffers
Prepare gels and assemble

electrophoresis cell
Prepare and load samples
Perform electrophoresis
Activate/image gels
Analyze the separation

Electrophoresis with Stain-Free Gels
Electrophoresis with Stain-Free Gels

Stain-free gels are made and packaged without sodium dodecyl sulfate (SDS),
allowing them to be used for both SDS and native polyacrylamide gel
electrophoresis (PAGE) applications.
To perform electrophoresis with stain-free gels

1. Prepare the sample and running buffers.
2. Set up the electrophoresis cell.
3. Perform the run.
Imaging Gels
Use unstained standards with stain-free gels, as some prestained standards are not
compatible with stain-free technology. To monitor electrophoresis, use a 1:1 mixture
of unstained and prestained standards.
Setting up a protocol for stain-free gels is similar to setting up protocols for other
applications. Follow the instructions in Creating a Protocol on page 20. Choose one
of the following activation times based on the sample and the purpose of your
experiment:
 Gels used in blotting — use 1 min activation for optimal results when
performing western blotting followed by immunodetection.
 Good sensitivity — use 2.5 min activation when samples are abundant
and when a fully optimized signal-to-noise ratio is not necessary.
 Best sensitivity — use 5.0 min activation for detection of proteins that are
in low concentration and for the best quantitation of the maximum number
of bands. Because the reaction is near completion after 5 min, this method
offers an optimal signal-to-noise ratio.
Note: If the gel has been activated for 2.5 min, activating it for another 2.5 min
might improve it. But activating an image for more than 5 min will not.
D | Using Bio-Rad Stain-Free Technology
Imaging Blots
To blot stain-free gels, use standard blotting procedures as described in the
instruction manual you are using. Use only PVDF membranes with low background
fluorescence, as membranes other than low fluorescence PVDF can result in high
background or low sensitivity with the imager.
To assess transfer efficiency, be sure to activate and visualize the gel using the
imager before transfer.

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ChemiDoc™ MP Imaging System

with Image Lab Software

Safety and Regulatory Compliance. . . . . . . . . . . . . . . . . . . . . . . . . . . .7

Chapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13

Instrument Guide | iii

Chapter 2 Acquiring Images. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

Chapter 3 System Calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

iv | ChemiDoc MP Imaging System with Image Lab Software

Epi Light Modules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51

Appendix D Using Bio-Rad Stain-Free Technology. . . . . . . . . . . . . 63

vi | ChemiDoc MP Imaging System with Image Lab Software

Important Safety Information

Use of the ChemiDoc MP imaging system involves UV illumination. Proper

 Defects caused by improper operation

 Repair or modification done by anyone other than Bio-Rad Laboratories,

 Use of spare parts supplied by anyone other than Bio-Rad Laboratories,

 Damage caused by accident or misuse

 Damage caused by disaster

 Corrosion caused by improper solvents or samples

 The instrument must be connected to a grounded power source line and

 Do not pour liquids directly on or inside the instrument.

 Switch off all lights on the instrument immediately after use.

 Clean the transilluminator sample area after use.

8 | ChemiDoc MP Imaging System with Image Lab Software

 Void the manufacturer’s warranty

 Void the regulatory certifications

 Create a potential safety hazard

Table 1. How alert icons are used in this guide

Extreme heat and flammable materials

10 | ChemiDoc MP Imaging System with Image Lab Software

Instrument Safety Warnings

Table 2. Safety cautions and warnings for the instrument

Warning: This instrument must be connected to an appropriate AC voltage

Power Safety Information

Voltage Setting Information

12 | ChemiDoc MP Imaging System with Image Lab Software

The ChemiDoc™ MP imaging system contains a charge-coupled device (CCD)

The imaging system includes the ChemiDoc MP instrument connected to a

 Control the ChemiDoc MP during image capture

 Optimize acquisition applications

 Produce reports and other output

ChemiDoc MP Imaging System

 Smart, application-based protocol setup using Image Lab software, which

 Exceptional sensitivity and a dynamic range greater than four orders of

 Flexibility to image chemiluminescent, fluorescent, and colorimetric

CCD Camera and Lenses

* U.S. patent 8913127

14 | ChemiDoc MP Imaging System with Image Lab Software

Universal Hood III

Image Lab Software

XcitaBlue Conversion Screen

Optional Light Sources

Green LED Module

Blue LED Module

16 | ChemiDoc MP Imaging System with Image Lab Software

18 | ChemiDoc MP Imaging System with Image Lab Software

Dynamic flat fielding Application-specific, for all applications

1. Select a protocol or customize a new one.

2. Position the gel or blot to be imaged.

3. Run your selected protocol.