Confocal Laser Scanning Microscope

FV3000
FLUOVIEW

Next Generation FLUOVIEW for the Next Revolutions in Science

 ( 1) (2)

(3)

1
The FLUOVIEW FV3000 Series − FV3000 and FV3000RS

The Next Evolution of Confocal Laser Scanning Microscope Technology for

Cell Biology, Cancer Research, Stem Cell Research, and Advanced Applications
The FLUOVIEW FV3000 Series is designed to meet some of the most difficult challenges in modern science. With
the high sensitivity and speed required for live cell and tissue imaging and the ease of use and flexibility required for
microplate imaging and complex screening protocols, the FV3000 Series supports complete workflows from live cell
2D–6D (x,y,λ,z,t,p) imaging through image processing, like deconvolution, and analysis. Particular attention has been
paid to the needs of cell biology (pages 5–6), cancer research (pages 7–8), and stem cell research (page 9). The
FV3000 is optimized for macro to micro imaging of cells, tissues, and small organisms.

With Olympus’ renowned optics at the heart of the system, the FV3000 features a new spectral detection concept for
true multichannel spectral imaging with high sensitivity detection in multiple dynamic ranges so even dim signals can be
separated. The optical path enables macro to micro imaging from 1.25X to 150X magnification combined with robust,
intuitive automation to simplify complex experiments, including one-click cellSens macro analysis for cell counting and
segmentation analysis. The precision of galvanometer scanning is combined with the speed of resonant scanning in the
FV3000RS hybrid scanner so users can combine precision and high-speed imaging in one experiment.

Built for long service life and low operating costs, the FV3000 uses long-lasting all diode lasers and LED illumination.
The system features a modular, upgradable design that includes 2-tier detection options, easily upgradeable laser
configurations, and the stable and flexible IX83 microscope with a field-upgradable z drift compensator (IX3-ZDC2)
for fast and robust live cell autofocus. With user-savable and selectable software workflows, the system adjusts to
individual needs. The facility manager tracking software makes it easy to track system usage by user, making the
FV3000 the ideal confocal system for years of productive science in single and multi-user environments.

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 The FV3000 Series: Meeting the Challenges of Cell Biology, Cancer
Research, Stem Cell Research, and Advanced Applications

Cell Division, Proliferation, Counting, Cell Cycle, and Microfluidics and High-Speed Blood Flow
Segmentation Analysis Circulating tumor cells in peripheral blood and microfluidic
Cell proliferation is a key aspect of cancer research. The FV3000 device imaging can require high-speed imaging for accurate
has tools for imaging and measuring these critical events. measurements. The FV3000RS provides high-speed imaging
(1) (2) (3) (4) for critical velocity measurements to capture key events.
(8)

Silicone Objectives Optimized for Live Tissue Observation

3D imaging has become an increasingly important part of
cancer research. Olympus’ exclusive silicone objectives provide
clear and bright images at depth in live cells and tissues for
accurate imaging and quantification.

5 μm 5 μm
Fast Calcium Dynamics
Image calcium sparks and waves at speeds up to 438 frames per
35 μm 35 μm second. Slow heartbeats to visible rates and capture vast neuronal
cell networks at full field of view at 30 frames per second.
UPLSAPO60XS2 UPLSAPO 60XO
(NA 1.3, W.D. 0.3 mm, silicone oil ne = 1.4)
(5 ) (NA 1.35, W.D. 0.15 mm, immersion oil ne = 1.52)
(6)
( 9) ( 10) ( 11) (12)

Macro to Micro and Whole Slide Imaging

Cell biology research demands the flexibility to image small
organisms at the macro scale down to the micro at high resolution.
The FV3000 Series features optics that enable macro to micro (13)
imaging for enhanced flexibility.

(7 )

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Spheroid, Gel Matrix, Long-Term Time-Lapse, and Super Resolution
Microplate Imaging Olympus’ patented* confocal super resolution imaging provides
Long-term time-lapse imaging of live cells in 3D captures an easy-to-use method for boosting resolution beyond the
physiologically relevant information. As stem cells grow into diffraction limit in fixed tissues.
spheroids and organoids, the FV3000 Series enables precise, *US8933418B/JP5784393B
stable time-lapse imaging with high sensitivity and low
FV-OSR
phototoxicity.

CONVENTIONAL CONFOCAL
( 18)
( 14 ) (15)

Photoconversion and Stimulation

(1 6 ) Precise control of laser light stimulation timing and complex
multipoint imaging and stimulation enable highly reproducible
experiments for various studies.

Spectral Unmixing
1050.0

Complex overlapping fluorescent protein spectra can complicate 1000.0

950.0
900.0
850.0

a range of biological studies. The FV3000 Series efficiently 800.0

750.0
700.0
650.0

separates signals for accurate measurements and localization. 600.0

Intensity

550.0
500.0
450.0
400.0
350.0
300.0

(1 7 ) 250.0
200.0
150.0
100.0
50.0

0 1000 2000 3000 4000 5000 6000 7000 8000 9000 100001100012000
Time(μs)

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 Solutions for Cell Biology: Image Dynamic in vivo Processes in Large
and Small Organisms with Very Low to High Magnification

Macro to Micro and Whole Slide Imaging

Cell biology requires high sensitivity, and deals with live organisms such as zebrafish and C. elegans. Large pieces of tissue and small
organisms may require both high speeds as well as large fields of view to see the entire organism in context. Accurately imaging a
large field of view requires precise automation and excellent optics. The FV3000 System is designed to image large tissues and small
organisms with accurate stage control, image stitching, and an optical design that facilitates very low to high magnification (1.25X up
to 150X). Since autofluorescence can be an issue for cell biologists, the FV3000 was designed to be a fully spectral system capable of
highly sensitive and accurate spectral background, autofluorescence, and overlapping spectra (e.g. GFP/YFP) separation.

1.25X Objective Single Shot Acquisition with Blind Unmixing

1.25X Objective Single Shot Acquisition

Mouse brain hemisection embedded for Expansion Microscopy (pre-expansion). Secondary antibody labels against GFP (Alexa Fluor 488, neurons), SV2 (Alexa
Fluor 565, Red) Homer (Alexa Fluor 647, Blue). Sample courtesy of Dr. Ed Boyden and Dr. Fei Chen, MIT.

Dendrite (anti-GFP Alexa Fluor 488, green) and synaptic

marker (SV2, Alexa Fluor 565, red) Olympus Super Resolution
image processed with cellSens advanced contrained
iterative deconvolution. Average Full Width Half Maximum
measurements ~135 nm. Image acquired with 100X 1.35 NA
silicone objective.
Sample courtesy of Dr. Ed Boyden and Dr. Fei Chen, MIT.

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A new optical design means that even when using low magnification 30X silicone objectives with 1.05 NA, resolution can be boosted
using Olympus super resolution technology—FV-OSR. Silicone objectives also help provide low spherical aberration on tissues and
small organisms, so object measurements and distances are accurate. The resonant scanner also helps reduce phototoxicity and
photobleaching compared to regular galvo scanners by reducing triplet states of excited fluorophores and reactive oxygen species.

Highly Dynamic Imaging

Small organisms are often favored as models for studying dynamic in vivo processes, so the FV3000RS is equipped with a very accurate
resonant scanner, facilitating applications such as studying a beating heart, blood flow, calcium signaling, and other dynamic events
at up to 438 frames per second. With the FV3000RS, switching between the high-precision galvanometer and high-speed resonance
scanner is as simple as a mouse click. The resonance scanner maintains the same field of view so users won't get lost when switching
between high-speed and high-precision scanning. Resonance images undergo post-processing with rolling average filtering for time gate
image averaging while improving signal-to-noise. Ratio imaging can employ an Intensity Modulated Display (IMD) so real signal stands
out above background noise. Selecting the spectral range is simple, and spectral unmixing is fast and automated.

Intensity Modulated Display of CFP/YFP ratio result during spontaneous contractions of in vitro cardiomyocyte.
Image data courtesy of Yusuke Nino and Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN.

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 Solutions for Cancer Research: Accurate 3D Cell and Tissue Imaging,
High-Speed Blood Flow, Microfluidic Imaging, and Robust Analysis

Cell Division, Proliferation, Counting, Cell Cycle, and Segmentation Analysis

The FV3000 Series incorporates the range of technologies necessary for cancer research imaging studies. In live cell cancer studies,
sensitive fluorescence detection, optimized optics, and analytical tools such as cell counting and segmentation analysis are essential.
With the emergence of microfluidics and a focus on circulating tumor cells, high-speed acquisitions can make the difference between
success and failure in an experiment.

Accuracy and repeatability are equally important; cell cycle checkpoint times must be reliably tracked, 3D images of cells must correctly
represent their shape and size, and images need to be bright and clear for segmentation analysis. Olympus’ silicone objectives are
optimized for tissue imaging. The FV3000 Series high-sensitivity cooled GaAsP detection unit with high signal-to-noise galvo and
resonant scanning and robust software make imaging accurate and reproducible for reliable results.

Platelets bound to thrombosis in blood vessel of mouse. Images taken 30 fps in full frame by resonant scanner with 2 CH GaAsP PMTs.
Image data courtesy of Dr. Takuya Hiratsuka, Dr. Michiyuki Matsuda, Graduate School of Biostudies, Kyoto University.

NK-cell mediated cell killing after therapeutic anitbody application (blue). GFP labeled NK-cells (green). DAPI uptake marking dead cells (Red).
Image data courtesy of Dr. Yuji Mishima, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research.

5 μm 5 μm

35 μm 35 μm

UPLSAPO60XS2 UPLSAPO 60XO

(NA 1.3, W.D. 0.3 mm, silicone oil ne = 1.4) (NA 1.35, W.D. 0.15 mm, immersion oil ne = 1.52)

Scal eA2-treated neocortex

Image data courtesy of Motokazu Uchigashima, M.D., Ph.D., Masahiko Watanabe, M.D., Ph.D., Departments of Anatomy, Hokkaido University Graduate School of Medicine.

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The system’s sensitivity coupled with the laser power monitor and two freely selectable ranges for laser power help provide that
apoptosis is part of the experiment and not caused by phototoxicity. The spectral sensitivity and accuracy enable researchers to conduct
multi-color fluorescence labeling experiments with multiple biomarkers.

Complex Tasks Made Simple

Cancer research is complex but measuring proliferation with the FV3000 isn't. With cellSens macro capabilities, time-lapse images can
be processed and counted and reports generated with a single mouse click. The layout of the acquisition software can be customized
according to specific applications and immediately selected on startup, making workflows logical and tailored to a customer's needs.
Specific experiment conditions can easily be reloaded, taking the guess work out of reproducing results.

Fucci cell cycle counting and expansion by cellSens.

Image data courtesy of Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN.

3D Time-lapse of mouse embryonic fibroblast labeled with silicone rhodamine docetaxol (Tubulin), imaged with 100X silicone objective and 30 fps resonant
scanning followed by cellSens deconvolution. Image data courtesy of Dr. Markus Delling, Harvard University.

A Scanning
size

B
Interval
time
0 min 30 min 60 min 90 min
Sequence Manager allows for variable time-lapse

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 Solutions for Stem Cell Imaging: Z Drift Compensator, and Intuitive Software
for Accurate Long-Term and Multipoint Time-Lapse Imaging in Microplates

Stem cell imaging requires increased levels of automation and long-term time-lapse capabilities. The FV3000 is designed to image
cells over multiple days with accurate timing, low phototoxicity, and accurate focus. Multipoint time-lapse in microplates is routine in
stem cell imaging, so the FV3000 can be enhanced with the IX3-ZDC2, Z drift compensator. The IX3-ZDC2 is designed to work with
the well navigator, so each well stays in focus during an experiment. For long experiments, add the laser power monitor to maintain
consistent laser exposure for excellent laser stability.

Users performing stem cell imaging benefit from high-sensitivity detection, silicone objectives, low phototoxicity from the resonant
scanner, and the higher throughput from high-speed scanning. Precise stimulation control means photoconversion is simple and
efficient, so cells can be reliably stimulated and imaged over multiple days for cell lineage tracking. Whether stem cell cultures are
in microplates, single dishes, or microfluidic devices, the FV3000 software and automation makes workflows simple. The stage
navigator includes well plate navigation and makes it easy to save, modify, and re-load frequently used plate settings and acquisition
conditions. Users can quickly image individual lanes of microfluidic channels. The sequence manager makes it easy to set up long-
term time-lapse imaging. Users can adjust the speed and timing of acquisitions while maintaining accurate timing. Quickly visualize
and download publication and presentation-ready 3D and 4D image data with the intuitive rendering software included with the
FV3000 software suite. Once imaging is completed, the macro functionality in cellSens analysis facilitates 2D cell counting and
segmentation with a single mouse click.

A spheroid image of a NMuMG cell line expressing Multipoint time-lapse window IX3-ZDC2
Fucci2.
Image data courtesy of Atsushi Miyawaki, Cell
Function Dynamics, Brain Science Institute of RIKEN.

MatTek EpiDermFT Tissue

Model: Immunofluorescence
labeled with 6 targets of
interest. 1. Abcam DRAQ5
ab108410, 2. Abcam Anti-
GAPDH (Alexa Fluor 405)
ab206372, 3. Abcam Anti-
Tubulin (Alexa Fluor 488)
ab1955883, 4. Abcam Anti-
Fibrillarin (Alexa Fluor 568)
ab202540, 5. Abcam Anti-
Vimentin (Alexa Fluor 594)
ab154207, 6. Abcam Anti-
Ki67 (Alexa Fluor 647) ab
194724. sample courtesy of
MatTek.

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Solutions for Advanced Applications: Spectral Unmixing, Super Resolution,
and Photostimulation

Both the FV3000 and FV3000RS have a range of standard and optional advanced application features including Olympus Super
Resolution (FV-OSR), photostimulation, spectral unmixing, and an external beam combiner. With precise laser control and Olympus’
patented super resolution method, the FV3000 Series can acquire images with a resolution down to 120 nm, similar to structured
illumination methods. Spectral unmixing is robust for a range of applications while photoconversion and photostimulation are efficient
and precise, enabling high-speed targeted path scanning and stimulation mapping studies.

The Sequence Manager makes it easy to reliably achieve complex cell cycle imaging protocols. Advanced applications, such as random
access or targeted path scanning, enable high signal-to-noise multipoint fluorescence measurements for in vitro neuronal cell signaling
studies while real-time processing and triggering help provide accurate and coordinated timing control for TTL-driven perfusion devices,
stimulators, or other 3rd party peripherals. Macro to micro functionality is easy with the FV3000 Series thanks to the stage navigator,
automation built into the IX83 microscope, and the ability to save and reload software layouts, workflows, and experiment conditions.

Spectral Umixing

Brainbow AAV transfection of Purkinje cells, amplified with antibodies as described in Cai et al 2013. Visible are Purkinje cell somata, dendrites and axons, as
well as some aspecific stainings of granule cells.

Photoconversion and Stimulation Super Resolution

1050.0
1000.0
950.0
900.0
CONVENTIONAL CONFOCAL
850.0
800.0
750.0
700.0
650.0
600.0
Intensity

550.0
500.0
450.0
400.0
350.0
300.0
250.0 FV-OSR
200.0
150.0
100.0
50.0

0 1000 2000 3000 4000 5000 6000 7000 8000 9000 100001100012000
Time(μs)

Trachea multi-ciliated epithelial cells (Culture)

Immunofluorescence microscopy: Odf2 staining (Alexa Fluor 488, green) of
cilia at the upper part of the basal body (green). Staining for ZO-1 revealed
the tight junctions (magenta).
Objective: UPLSAPO60XS
Image data courtesy of Hatsuho Kanoh, Elisa Herawati, Sachiko Tsukita,Ph.
D. Graduate School of Frontier Biosciences and Graduate School of
Medicine, Osaka University.

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 FV3000 with Galvanometer Scanner to FV3000RS with Resonant
Hybrid Scanner: Flexible Configurations to Advance Science

High-Speed Resonant Scanning up

to 438 Frames per Second
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Flexible Detection Lightpath

with Wide Dynamic Range
Photomultiplier Tubes (PMTs) or
High Signal-to-Noise, Cooled
GaAsP Spectral Detection Concept
(2–4 Channels)
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Multichannel Spectral Detector

with 16-Channel Unmixing
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Combiner System Featuring

Diode Lasers with a Range of
Wavelengths
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Advanced Olympus Optics

Page
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Z Drift Compensator—IX3-ZDC2
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Precise Ultrasonic Stage IX3-SSU

for Multi-Area Imaging
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No Darkroom Required
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Powerful, Intuitive Software
15
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g

Precise Sequence Manager and

Real-Time Acquisition
17
Page
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Well Navigator for Microplate,

Multipoint Time-Lapse Imaging,
and Stitching
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Powerful One-Click cellSens Macro

Analysis
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Olympus Super Resolution with Up

to 4 Simultaneous Channels
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FV3000RS
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 The Right Mixture of Speed and Accuracy

The FV3000 Series Scan Units

Galvanometer and Galvo/Resonant Hybrid Scanner Optimized for Live Cell Imaging
Users have their choice of two different types of scan Resonant scanning greatly reduces photobleaching and
units: galvanometer only with the FV3000 or galvanometer/ phototoxicity compared to standard galvanometer scans by
resonant hybrid with the FV3000RS. The hybrid scan unit has preventing the excitation of fluorophores into triplet states that
galvanometer scanners for high-precision scanning, as well create reactive oxygen species. These features make live cell
as a galvo/resonant scanner ideal for high-speed imaging. experiments more robust and reliable. The FV3000 Series has
Galvanometer scanner enables Olympus super resolution complete high and low range laser intensity control enabling the
technology (FV-OSR) yields resolutions down to 120 nm as well system to use the minimum required amount of laser power on
as high signal-to-noise, with precise tornado and multipoint samples. The optional Laser Power Monitor provides consistent
stimulation and 100 ms switching time. Galvanometer scanning laser power during long-term time-lapse imaging across multiple
can achieve 16 frames per second at 2X zoom. The resonant days.
scanner is capable of speeds ranging from 30 frames per second
at 512 × 512 to 438 frames per second at 512 × 32.

No Compromise between Speed and Field of View

Many high-speed scanning methods restrict the field of view,
limiting their usefulness for examining large areas with multiple
cells. The FV3000 Series’ resonant scanner maintains a full
1X field of view, even at a video rate of 30 frames per second.
Additional speed is generated by clipping the Y axis, even at 438
frames per second.

Platelets bound to thrombosis in blood vessel of mouse. Images taken 30

fps in full frame by resonant scanner with 2 CH GaAsP PMTs.
Image data courtesy of Dr. Takuya Hiratsuka, Dr. Michiyuki Matsuda,
Graduate School of Biostudies, Kyoto University.

Other
common
scanners

FV3000
FN 18, 8 KHz

Most resonant scanners force a trade-off between speed and

field of view. FLUOVIEW systems are optimized to maintain the
field of view with even signal intensity so dynamic samples (e.g.
calcium imaging) can be seen in the broad context of their cells
and tissues.
The image above shows examples of the zoom factors required
in other systems.

A431 cells fixed with methanol labeled with Abcam Anti-ERK1 + ERK2
antibody (Alexa Fluor 488) ab208564, and Anti-alpha Tubulin antibody (Alexa
Fluor 594) ab195889 and DAPI.
Sample courtesy of Abcam.

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Introducing TruSpectral Detection

A Fully Spectral System with Sensitivity and Accuracy Efficient TruSpectral Detection System
The FV3000 Series employs Olympus’ TruSpectral detection The FV3000 Series brings new levels of total system transmission
concept. Based on patented* Volume Phase Hologram (VPH) efficiency, enabling every system to be completely spectral,
transmission and an adjustable slit to control light, the spectral improving overall sensitivity, and improving the signal-to-noise
detection in FV3000 and FV3000RS is highly efficient, enabling ratio for improved multi-color confocal imaging.
users to select the detection wavelength of each individual Transmission Efficiency Ratio of FV3000 to FV1200
(Normalization FV1200 as 1.0)
channel to 2 nm. 1.5
* US8530824B/JP5541972B/EP2395380A FV3000
1.4
FV1200

Efficiency ratio
Tube Lens 1.3

1.2

1.1

1
Adjustable slit
0.9
400 450 500 550 600 650
Wavelegnth (nm)

Multichannel TruSpectral Detection with 16-Channel

VPH
Unmixing
TruSpectral’s efficient design and software enable spectral
detectors to run in multichannel mode for both live and post-
processing spectral unmixing with a multichannel lambda
mode. Multichannel mode facilitates constant spectral
High-Sensitivity Spectral Detector (HSD) with GaAsP unmixing during live cell experiments, separating complex
Photomultiplier Tubes Enhances Quantum Efficiency fluorescence during acquisition. With up to 4 different dynamic
HSD makes it possible to view samples that were too dim to ranges from the 4 different channels of array, even bright
view with conventional equipment. The GaAsP PMT incorporates and dim spectral signals can be separated by adjusting the
2 channels with a maximum quantum efficiency of 45 %, and sensitivity of each detector independently.
Peltier cooling reduces background noise by 20 % for high S/N
ratio images under exceptionally low excitation light.

Standard Quantum Efficiencies of Detector Technologies

400 500 600 700

40
Sensitivity Adjustment of Each Channel
Quantum Efficiency (%)

30 GaAsP PMT

20

400 500 600 700

10 Multi-Alkali PMT Spectral Unmixing

0
400 500 600 700 800 900
Wavelength (nm)

400 500 600 700 400 500 600 700

400 500 600 700 400 500 600 700

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 From Basic to Advanced Acquisition and Analysis, an Interface
that Adapts to Your Workflow

Intuitive Workflow 1. Layout 2. Acquisition 3. Acquisition

Customizable and saveable layouts Start by selecting Condition Activate basic to complex acquisitions
make it easy to tailor the interface to your preferred Reload settings that with live ratio, intensity modulated display,
your workflow and experiment needs, display with specific were ideal for your quantitative region of interest (ROI)
from basic to complex. tools for basic to last experiment to graphing or spectral unmixing display, and
complex acquisition. provide consistency. data backup for added security.

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4.Viewer 5. Analysis
Live Spectral Unmixing with
Review data as it is Extract data from images using online or offline TruSpectral Detection
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generated. Generate processing. Analytical tools include Olympus super
3D and 4D views and resolution technology (FV-OSR) and powerful cellSens
animations to explore and software with features such as deconvolution, filtering,
share data in depth. count and measure, and one-click macros. Sequence Manager
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Stage Control for Multipoint

Time-Lapse and Microplate
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Hard Drive Data Backup

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One-Click Macro Analysis

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Ratio Imaging and Intensity

Modulated Display (IMD)
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Rolling Average Processing

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Deconvolution
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FV-OSR (Olympus Super

Resolution) Technology
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Macro to Micro Observation

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A431 cells fixed with 100 % methanol. Abcam Anti-

Integrin alpha 2 antibody (Alexa Fluor 488) ab208770,
and Anti-alpha Tubulin antibody (Alexa Fluor 594)
ab195889 and DAPI.
Sample courtesy of Abcam.

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 Intuitive Stage Control, Live Spectral Unmixing, Real-Time Acquisition

Live Spectral Unmixing with TruSpectral Detection and Stage Control for Multi-Area Time-Lapse, Microplate,
Real-Time Processing and Stitching
The power of TruSpectral detection plus multichannel mode Microplate imaging is important for many applications, and the
means live spectral unmixing can be performed during Well Navigator provides sophisticated, intuitive controls for a wide
image acquisition, providing real-time processing of complex range of cell culture vessels and custom plates. Multi-area time-
overlapping spectra. lapse and stitching provide robust and accurate time-lapse data.

Live blind unmixing of CFP (endosomes, blue), mAmetrine (plasma membrane, green),
mKO (nucleus, orange) and mKeima (F-actin, purple) during time-lapse imaging.
Image data courtesy of Dr. Kazuhiro Aoki, Dr. Michiyuki Matsuda, Graduate School of
Medicine, Kyoto University.

Maintain Focus with Z Drift Compensation (ZDC) System

The IX3-ZDC2 uses minimally-phototoxic IR light (laser class 1) Hard Disk Recording
to identify the location of the sample plane. One-shot autofocus The microscope comes equipped with a hard-disk drive
(AF) mode allows several focus positions to be set as desired for (HDD) recording function. The images captured are stored
deeper samples, enabling efficient Z-stack acquisitions in multi automatically in the HDD. Large volumes of data, such as those
position experiments. Continuous AF mode keeps the desired obtained from long-term time-lapse imaging can be stored.
plane of observation precisely in focus, avoiding focus drift due
to temperature changes or the addition of reagents, making it
ideal for measurements that requires more stringent focusing. Powerful One-Click Macro Analysis with cellSens
Furthermore, increased optical offset enables continuous AF Images alone are not enough; with integrated cellSens Count
over plastic vessels or with dry objectives. The IX3-ZDC2 is also and Measure analysis, the FV3000 Series can optimize images
compatible with silicone objectives (in AF mode). with deconvolution and analyze them with one-click macro
cell functionality for a broad range of morphological measurements.
IX3-ZDC2 Optical Path Diagram
Offset

Offset lens
Cover glass
Objective
AF sensor

Precise Sequence Manager and Real-Time Acquisition

Complex protocols are handled with ease, and real-time control A spheroid image of a NMuMG cell line expressing Fucci2.
Image data courtesy of Atsushi Miyawaki, Cell Function Dynamics, Brain
helps provide microsecond accuracy of scans with millisecond Science Institute of RIKEN.
accuracy over days of time-lapse.

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Additional Intuitive Features

Ratio Imaging and Intensity Modulated Display (IMD) Rolling Average Processing
The FV3000RS includes an Intensity Modulated Display (IMD) High-speed scanning at low laser power to avoid phototoxicity
function in the software that displays quantitative fluorescence often decreases the signal-to-noise ratio. With rolling average
ratio changes during both standard and high-speed acquisitions. post-processing, users have the flexibility to adjust high-speed
This function is particularly useful for calcium and FRET imaging time-lapse images while maintaining time scale and keeping the
where a pure ratio display provides poor contrast in background original data.
areas.

Ratio(ex405/ex488)

High

Raw 30 fps data acquired at low Rolling average processing (10

laser power (0.05 %, 488 nm). frame) on 30 fps data acquired at
low laser power.

Low
10 μM CCCP treatment
Deconvolution
Ratio(ex405/ex488)
The optional Constrained Iterative (CI) Deconvolution Solution
employs advanced CI algorithms to produce improved resolution,
contrast, and dynamic range, with industry-leading speed.
This proprietary post-processing tool is efficient for both CCD
and confocal imaging and enhances the ability to differentiate
between imaged objects.

tsGFP1-mito reveals heterogeneity in mitochondrial thermogenesis in HeLa cells.

The images of ratio (ex 405 nm/ex 488 nm) in tsGFP1-mito-expressing cells
before and after CCCP treatment at 37 °C. Scale bars indicate 10 μm (whole
image) and 3 μm (inset).
Image data courtesy of Shigeki Kiyonaka Ph,D, Yasuo Mori Ph,D Molecular
Biology Field, Department of Synthetic Chemistry and Biological Chemistry,
Kyoto University.

Original Image Deconvolved Image

Cell line: Human cervical cancer cell line HeLa cell

Immunostaining: Hec1 staining (green, Alexa Fluor 488), α-tubulin staining
(red, Alexa Fluor 568),DAPI staining (blue)
Mitotic HeLa cell derived from human cervical cancer.
Mitotic spindle and kinetochores are stained with anti-α-tubulin (red) and
anti-Hec1 (green) antibodies, respectively. Chromosomes interact with
CFP YFP FRET
microtubules constituting mitotic spindle via kinetochores, protein structure
assembled on centromere region of chromosomes.
Image data courtesy of Masanori Ikeda and Kozo Tanaka, Department of
Molecular Oncology, Institute of Development, Aging and Cancer, Tohoku
University.

Raw CFP/YFP ratio IMD of CFP/YFP ratio

Cardiomyoctye
Image data courtesy of Yusuke Niino and Atsushi Miyawaki, Cell Function
Dynamics, Brain Science Institute of RIKEN.

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 Olympus Super Resolution Technology

Olympus Super Resolution (FV-OSR) Technology with Up to 4 Simultaneous Channels

Olympus’ widely applicable super resolution method requires no special fluorophores and works for a wide range of samples. Ideal
for colocalization analysis, the FV-OSR can acquire 4 fluorescent signals either sequentially or simultaneously with a resolution of
approximately 120 nm*, nearly doubling the resolution of typical confocal microscopy. The system is easy to use with minimal user
training and can be added to any confocal system, making the FV-OSR a truly accessible method for achieving super resolution.
* Subject to objective magnification, numerical aperture, excitation and emission wavelength, and experiment conditions.

Beyond Deconvolving Confocal: Comparison of Confocal, Deconvolved Confocal and Deconvolved FV-OSR Images
0.5 AU Confocal Image Enlargement

0.5 AU Confocal Image Deconvolved with cellSens Advanced Deconvolution

Olympus Super Resolution Plus cellSens Advanced Deconvolution

Secondary antibody labels against GFP (Alexa Fluor 488,

neurons) and SV2 (Alexa Fluor 565, red).
Sample courtesy of Dr. Ed Boyden and Dr. Fei Chen, MIT.

Macro to Micro Observation

Finding areas of interest in samples can be challenging. The confocal optical design of the FV3000 Series supports macro to micro
imaging so users can quickly switch from low magnification overview observation with 1.25X objectives to high-magnification, detailed
observation with up to 150X objectives. Users can employ image stitching at both macro and micro levels to generate overview
images that show samples in context.

A stitched image of a coronal section (30

μm thickness) from an adult YFP-H mouse
cerebrum acquired with 20X objective
(UPLSAPO20X).
Image data courtesy of Takako Kogure and
Atsushi Miyawaki, Cell Function Dynamics,
Brain Science Institute of RIKEN.

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Superior Optics and a Rigid Frame Ideal for Live Cell Imaging

Silicone Immersion Objectives for Live Cell Imaging Deliver High-Resolution Observation at Depth
Olympus offers four high NA silicone immersion objectives that deliver excellent performance for live cell imaging. The refractive index of
silicone oil (ne≈1.40) is close to that of living tissue (ne≈1.38), enabling high-resolution observations deep inside living tissue with minimal
spherical aberration caused by refractive index mismatch. Silicone oil does not dry out or harden, so there is never a need to refill oil,
making it ideal for extended time-lapse observations.
Refractive Index is Important with Deep Tissue Observation
UPLSAPO30XS: For a broader view and greater depth In deep tissue observation, image quality depends on keeping the refractive index of the
Magnification: 30X, NA: 1.05 (silicone oil immersion), W.D.: 0.8 mm, sample and immersion medium as close to each other as possible.
cover glass thickness: 0.13–0.19 mm, operating temperature: 23 –37 °C

UPLSAPO40XS : Complete the magnification range

Magnification: 40X, NA: 1.25 (silicone oil immersion), W.D.: 0.3 mm, Sample
cover glass thickness: 0.13–0.19 mm, operating temperature: 23 –37 °C
ne≈1.38
UPLSAPO60XS2: For 3D observations with superior resolution
Magnification: 60X, NA: 1.30 (silicone oil immersion), W.D.: 0.3 mm, Water Cover glass Silicone oil
cover glass thickness: 0.15–0.19 mm, operating temperature: 23 –37 °C ne≈1.33 ne≈1.52 ne≈1.40
UPLSAPO100XS: For greater depth in closely defined regions
Magnification: 100X, NA: 1.35 (silicone oil immersion), W.D.: 0.2 mm, Water immersion objective Silicone immersion objective
cover glass thickness: 0.13–0.19 mm, operating temperature: 23 –37 °C When working with a water immersion When working with a silicone immersion
objective, the difference between the objective, the difference between the
refractive index of the samples and water refractive index of the samples and silicone
results in spherical aberration in deep oil is minimal. So it achieves brighter
tissue, causing resolution to deteriorate and fluorescence images with higher resolution
fluorescence to become dim. for deep tissue.

PLAPON60XOSC2: Enhance the Reliability of Colocalization Analysis with a Low Chromatic Aberration Objective
This oil immersion objective minimizes lateral and axial chromatic Performance Comparison of PLAPON60XOSC2 and UPLSAPO60XO
aberration in the 405–650 nm spectrum. Colocalization images PLAPON UPLSAPO
are acquired reliably and images are measured with superior 60XOSC2 60XO
positional accuracy. The objective also compensates for
Axial chromatic aberration (Z direction)
chromatic aberration through near infrared up to 850 nm, making Compared for PSF fluorescent beads Approx. Approx.
0 μm 0.5 μm
it the ideal choice for quantitative imaging. (405 nm, 633 nm)

Lateral chromatic aberration (X-Y direction)

Low Chromatic Aberration Objective Compared for PSF fluorescent beads
Magnification: 60X (405 nm, 488 nm, 633 nm) Approx. 0.1 μm Approx. 0.2 μm
NA: 1.4 (oil immersion)
W.D.: 0.12 mm
3D image
Chromatic aberration compensation range: Tubulin in Ptk2 cells labeled with two colors
405–650 nm (405 nm, 635 nm) and compared
Optical data provided for each objective.

Meeting the Requirements of Stability with the IX83 High Contrast under Bright Conditions
A Z-drive guide installed near the revolving nosepiece combines The umbra unit is designed specifically for fluorescence
high thermal rigidity with the stability of a wraparound structure to observation. It efficiently blocks out room light, enhances
significantly reduce the impact of heat and vibration and improve the contrast of fluorescence, and enables clear fluorescence
the quality of time-lapse imaging. observation under bright conditions.
Thermal Drift Displacement Periodic Damping

IX81

IX81

IX83
Square Frame for
Increased Rigidity
IX83

0 50 100 150 200 (min) 0.0 0.5 1.0 1.5 2.0 (s)

20
 Modular Units Designed for Your Applications

Scanners Laser Combiners Other Equipment

Choose from the following options with field-
upgradable laser-based autofocus, fast and
precise motorized stage control, analog
input/output and TTL synchronization, and a
convenient anti-vibration platform.

Hybrid Scan Unit (Resonant/Galvanometer) Main Laser Combiner

The hybrid scanner combines the capabilities The main laser combiner is the heart of the laser
of a galvanometer scanner with a resonant system. Four standard lasers with an option to
scanner for high-speed imaging in the full field add a fifth laser or leave an open port to add
of view at 30 fps and up to 438 fps at 512 × an additional three diode lasers via the Sub
32. The Sequence Manager makes it simple Combiner.
to automatically switch between resonant and Z Drift Compensator/IX3-ZDC2
galvanometer imaging in the same experiment. Sub Laser Combiner The IX3-ZDC2 uses minimally-phototoxic IR
Add this optional combiner at any time with up light to identify the location of the sample plane.
Galvo Scan Unit to 3 diode lasers for a maximum of 7 laser lines The IX3-ZDC2 is also compatible with silicone
The galvanometer-only scanner provides in combination with the main laser combiner. objectives and plastic bottom vessels.
precision scanning from 1 fps at 512 × 512
to 16 fps. High-speed multipoint stimulation
or detection experiments can travel between
multiple cells at over 100 Hz with data output
as high as 500 kHz.
Illumination Units
The conventional illumination modules
are designed for long-duration time-lapse
experiments. Since light is introduced through
fiber delivery systems, no heat is transferred to
Spectral Detectors the microscope.

Ultrasonic Stage for IX3/IX3-SSU

With low thermal drift for improved accuracy,
the ultrasonic stage can be controlled by both
software and Touch Panel Control for fast, reliable
multi-area imaging.

High Sensitivity Spectral Detector (GaAsP PMT) Light Source/U-HGLGPS

with TruSpectral Technology The pre-centered fluorescence illumination
The 2-channel High Sensitivity Spectral source requires no adjustment and has an
Detector (HSD) employs the same Volume average lifespan of 2,000 hours.
Phase Holographic (VPH) technology as the
SD, with Peltier cooled GaAsP PMTs and a high
quantum efficiency of 45 % and detection up
to 750 nm. This unit can be combined with the IO Interface Box/FV30-ANALOG
2-channel spectral detector (SD) for a flexible This unit supports electro-physiological
dynamic range or a second 2-channel HSD unit experiments through analog inputs and TTL I/
for powerful 4-channel sensitivity. O support. The interface box converts voltage
to images that can be treated in the same
Spectral Detector (Multialkali PMT) with manner as fluorescence images.
TruSpectral Technology
The 2-channel SD employs efficient VPH
transmission and an adjustable slit with 1–100
nm bandwidth from 400–800 nm detection. Transmitted Detector/FV31-LETD
The multialkalai PMTs provide a broad This unit combines an external transmitted light
dynamic range for detection up to 800 nm. photomultiplier detector and LED conventional
illumination for both laser scanning and
conventional transmitted light Nomarski DIC
observation. Users can undertake simultaneous
multichannel confocal fluorescence imaging
and transmitted DIC acquisition.

Simple Anti-Vibration Plate/FV31-AVP

Designed to match the footprint of the FV3000,
this simple anti-vibration plate provides a
compact solution for those who do not need a
full anti-vibration table.

21
Specifications

FLUOVIEW FV3000 Specifications

FV3000 FV3000RS
Laser Light Violet/Visible Light Laser 405 nm: 50 mW, 488 nm: 20 mW, 561 nm: 20 mW, 640 nm: 40 mW
One optional laser port for sub laser combiner or optional laser unit
Optional Laser Sub Laser Combiner Laser as follows (max. 3 laser units )
445 nm: 75 mW, 514 nm: 40 mW, 594 nm: 20 mW, connected to main laser combiner
Single Laser Unit 445 nm: 75 mW, 514 nm: 40 mW, or 594 nm: 20 mW, directly connected to main laser combiner
Laser Light Control Main laser combiner with implemented AOTF system, ultra-fast intensity modulation with individual laser lines, additional
shutter control continuously variable (0.1 %–100 %, 0.1 % increments)
10 % or 100 % maximum laser power changer by ND filter
Scanner Scanning Method 2 silver-coated galvanometer scanning mirrors 2 silver-coated galvanometer scanning mirrors
1 silver-coated resonant and 1 silver-coated galvanometer
scanning mirrors.
Galvanometer Scanner Scanning Resolution: 64 × 64 to 4096 × 4096 pixels
(Normal Imaging) Scanning Speed (One Way): 512 × 512 with 1.1 s — 264 s. pixel time : 2 μs — 1000 μs.
Scanning Speed (Round Trip): 512 × 512 with 63 ms - 250 ms
256 × 256 with 16 ms - 125 ms
Optical Zoom: 1X – 50X in 0.01X increments
Scan Rotation: Free rotation (360 degree) in steps of 0.1 degree
Scanning Mode: PT, XT, XZ, XY, XZT, XYT, XYZ, XYλ, XYZT, XYλT, XYλZ, XYλZT
ROI Scanning, rectangle clip, ellipse, polygon, free area, line, free line and point, tornado mode only for stimulation
Resonant Scanner Scanning Resolution: 512 × 32 to 512 × 512 pixels
(High-Speed Imaging) Scanning Speed: 30 fps at 512 × 512, 438 fps at 512 × 32
Optical Zoom: 1X – 8X in 0.01X increments
–
Scanning Mode: XT, XZ, XY, XZT, XYT, XYZ, XYλ, XYZT,
XYλT, XYZ, XYλZT
ROI Scanning, Rectangle Clip, Line
Pinhole Single motorized pinhole, pinhole diameter ø50 – 800 μm (1 μm Steps)
Field Number (FN) 18
Dichromatic Mirror Turret 8 positions (high performance DMs and 10/90 mirror)
Optional Unito for Scanner Laser Power monitor, optional laser port
High Sensitivity- Detector Module Cooled GaAsP photomultiplier, 2 channels
Spectral Spectral Method Motorized Volume Phase Holographic transmission diffraction grating, motorized adjustable slit,
Detector selectable wavelength bandwidth: 1–100 nm, wavelength resolution: 2 nm
Dichromatic Mirror Turret 8 positions (high performance DMs and mirror)
Spectral Detector Module Multi-Alkali photomultiplier, 2 channels
Detector Spectral Method Motorized Volume Phase Holographic transmission diffraction grating, motorized adjustable slit
selectable wavelength bandwidth: 1–100 nm, wavelength resolution: 2 nm
Dichromatic Mirror Turret 8 positions (high performance DMs and mirror)
Microscope Motorized Microscope Inverted IX83 (IX83P2ZF)
Integrated motorized focus module, minimum increment 0.01 μm
System Control Control Unit OS: Windows 7 Professional 64-bit (English version),
built-in dedicated I/F board and hardware sequencer for precise imaging timing
Display 30 or 32-inch monitor (WQUXGA 2560 × 1600)
Fluorescence Illumination Unit External fluorescence light source, fiber adapter to optical port of scan unit, motorized switching between LSM light path
and fluorescence illumination
Transmitted Light Detector Unit Module with integrated external transmitted light photomultiplier detector and LED lamp, motorized switching

Software
Basic Features GUI designed for darkroom environment. User-arrangeable layout.
Acquisition parameter reload features. Hard disk recording capability, adjust laser power and HV with Z-stack acquisition.
Z-stack with alpha blending, maximum-intensity projection, iso-surface rendering.
2D Image Display Each image display: single-channel side-by-side, merge, cropping, live tiling, live tile, series (Z/T/λ), LUT: individual color
setting, pseudo-color, comment: graphic and text input
3D Visualization and Observation Interactive volume rendering: volume rendering display, projection display, animation displayed.
3D animation (maximum intensity projection method, α blending) 3D and 2D sequential operation function
Image Format OIR image format
8/16-bit gray scale/index color, 24/ 32/ 48-bit color, JPEG/ BMP/ TIFF image functions, Olympus multi-tif format
Spectral Unmixing Fluorescence spectral unmixing modes (up to 16 channels)
Image Analysis Fluorescence intensity and time-lapse measurement
Statistical Processing 2D data histogram display
Optional Software Motorized-stage control
Mapping and multiplepoint simulation
Sequence manager
Virtual channel acquisition
Microplate navigation
Remote development kit
Super resolution imaging (FV-OSR)

World Wide Support

Installation generally takes one day to get systems up and running fast. We support our products via our global knowledge base.
Olympus application specialists can assist you with choosing the features that will optimize your system for your applications. Confocal
systems are an investment, and keeping the system running in the best performance is important. Our certified service teams can deploy
rapid alignment procedures and system diagnostics to keep your system in top shape and diagnose any issues.

22
 Image data are courtesy of the following institutions:

Mouse kidney (cover and page 2) and rat embryo sample (3, page 1) prepared
by Dr. Mike Davidson. Images presented with lasting gratitude for his lifetime
commitment to science and microscopy.

Whole mouse kidney captured in single shot with 1.25X objective. 10 μm section,
TOMM20 ATTO 647N, Phalloidin Alexa Fluor 568, WGA Alexa Fluor 488, DAPI.
(cover page and page 2)

3D rendered image of Xenopus endoderm labeled with malachite green and

methylene blue. 3 channel image captures label and autofluorescence. (1, page 1)

3D rendered image of Xenopus endoderm labeled with malachite green and

methylene blue. (2, page 1)

2 × 2 tiled image of whole rat embryo, 20 mm total field of view. H&E fluorescence
with 640 nm laser diode. (3, page 1)

Growing HeLa cells expresses Fucci, a cell cycle indicator. Fluorescense image (1,
page 3), Cell counting (2,3, page 3)
Asako Sakae-Sawano, Atsushi Miyawaki, Cell Function Dynamics, Brain Science
Institute of RIKEN.

3D Time-lapse of mouse embryonic fibroblast labeled with silicone rhodamine

docetaxol (Tubulin, white), RFP centrin (green) imaged with 100X silicone
objective and 30 fps resonant scanning followed by cellSens deconvolution. Dr.
Markus Delling, Harvard University. (4, page3)

Scal eA2-treated neocortex

Motokazu Uchigashima, M.D., Ph.D., Masahiko Watanabe, M.D., Ph.D.,
Departments of Anatomy, Hokkaido University Graduate School of Medicine. (5,6,
page 3)

A stitched image of a coronal section (30 μm thickness) from an adult YFP-H

mouse cerebrum acquired with 20X objective (UPLSAPO20X).
Takako Kogure and Atsushi Miyawaki, Cell Function Dynamics, Brain Science
Institute of RIKEN. (7, page3)

Platelets bound to thrombosis in blood vessel of mouse. Images taken 30 fps in

full frame by resonant scanner with 2 CH GaAsP PMTs.
Dr.Takuya Hiratsuka, Dr. Michiyuki Matsuda, Graduate School of Biostudies,
Kyoto University. (8, page 3)

FRET spectral look up table display of cardiac myocyte (9, page 3), CFP spectral
look up table display of cardiac myocyte (10, page 3) , Differential Interference
Contrast (DIC) image of cardiac myocyte (11, page 3), Overlay (12, page 3),
IMD ratio images of spontaneous Ca2+ oscillation in a beating rat cardiomyocyte
expressing yellow cameleon. (13, page 3)
Yusuke Niino and Atsushi Miyawaki, Cell Function Dynamics, Brain Science
Institute of RIKEN.

Fucci induced Spheroid of HT29 cell line

Yuji Mishima, Ph.D., Kiyohiko Hatake M.D., Ph.D. Clinical Chemotherapy, Cancer
Chemotherapy Center, Japanese Foundation for Cancer Research. (14, page 4)

A spheroid image of a NMuMG cell line expressing Fucci2.

Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN. (15,
page 4)

FRET imaging by expressed Raichu-Cdc42 in cultured HT1080. Activated Cdc42

is observed to the cell moving direction.
Ms. Satsuki Fujiwara and Dr. Michiyuki Matsuda, Graduate school of Biostudies,
Kyoto University. (16, page 4)

Brainbow AAV transfection of Purkinje cells, amplified with antibodies as

described in Cai et al 2013. Visible are Purkinje cell somata, dendrites and axons,
as well as some aspecific staining of granule cells. (17, page 4)

Cultured epithelial HeLa (EpH) cells.

Immunofluorescence microscopy: α-tubulin staining (Alexa Fluor 488, green),
ZO-1 staining (Alexa Fluor 568, magenta)
Staining for ZO-1 revealed the tight junctions (TJs) (magenta).
Staining for α-tubulin showed an apical network of microtubules. This network
associates with the TJ to form the “TJ-apical complex” (green).
Objective: UPLSAPO100XS
Image data Courtesy of
Hatsuho Kanoh, Tomoki Yano, Sachiko Tsukita,Ph.D. Graduate School of Frontier
Biosciences and Graduate School of Medicine, Osaka University. (18, page 4)

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