Confocal Laser Scanning Microscope
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, the FV3000 also provides an intuitive and adaptable interface, and is capable of microplate imaging and complex screening protocols. The 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, cancer research, and stem cell research. The FV3000 is optimized for macro to micro imaging of cells, tissues and small organisms.
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Cell proliferation is a key aspect of cancer research. The FV3000 has tools for imaging and measuring these critical events.
Loading the player… FV3000: Cell Segmentation Animation | Automated one click cellSens macro segmentation and analysis of Growing HeLa cells expresses Fucci, a cell cycle indicator.
Fluorescense image
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Loading the player… FV3000: Cell division resonant |
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.
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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.
ScaleA2-treated neocortex
Motokazu Uchigashima, M.D., Ph.D., Masahiko Watanabe, M.D., Ph.D., Departments of Anatomy, Hokkaido University Graduate School of Medicine.
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 imaging for enhanced flexibility.
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.
Circulating tumor cells in peripheral blood and microfluidic device imaging can require high-speed imaging for accurate measurements. The FV3000RS provides high-speed imaging for critical velocity measurements to capture key events.
Loading the player… FV3000: Blood Flow Video | 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) |
Image calcium sparks and waves at speeds up to 438 frames per second. Slow heartbeats to visible rates and capture vast neuronal cell networks at full field of view at 30 frames per second.
FRET spectral look up table display of cardiac myocyte (1), CFP spectral look up table display of cardiac myocyte (2), Differential Interference Contrast (DIC) image of cardiac myocyte (3), Overlay (4), IMD ratio images of spontaneous Ca2+ oscillation in a beating rat cardiomyocyte expressing yellow cameleon (5).
Yusuke Niino and Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN.
Long-term time-lapse imaging of live cells in 3D captures physiologically relevant information. As stem cells grow into spheroids and organoids, the FV3000 Series enables precise, stable time-lapse imaging with high sensitivity and low phototoxicity.
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.
Loading the player… FV3000: Spheroid animation | A spheroid image of a NMuMG cell line expressing Fucci2. Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN. |
Loading the player… FV3000: Fujiwara-crop | 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. |
Complex overlapping fluorescent protein spectra can complicate a range of biological studies. The FV3000 Series efficiently separates signals for accurate measurements and localization.
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.
Olympus’ patented* confocal super resolution imaging provides an easy-to-use method for boosting resolution beyond the diffraction limit in fixed tissues. *US8933418B/JP5784393B
Cultured epithelial HeLa (EpH) cells.
Immunofluorescence microscopy: a-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 a-tubulin showed an apical network of microtubules. This network associates with the TJ to form the “TJ-apical complex” (green).
Objective: UPLSAPO100XS
Hatsuho Kanoh, Tomoki Yano, Sachiko Tsukita,Ph.D. Graduate School of Frontier Biosciences and Graduate School of Medicine, Osaka University.
Precise control of laser light stimulation timing and complex multipoint imaging and stimulation enable highly reproducible experiments for various studies.
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.
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.
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. Multiple time point display of live cell. Quantitative graph result of ratio from ROI inside cell shown below.
Image data courtesy of Yusuke Nino and Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN.
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.
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.
Blood flow imaging and velocity measurements by cellSens software kymograph analysis.
Image data courtesy of Dr.Takuya Hiratsuka, Dr. Michiyuki Matsuda, Graduate school of Biostudies, Kyoto University.
Loading the player… FV3000: Dudi-158V-XYT-0001_00000 | NK-cell mediated cell killing after therapeutic anitbody application (blue). GFP labeled NK-cells (green). DAPI uptake marking dead cells (Red). Image data courtesy Dr. Yuji Mishima, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research. |
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 user'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. Multiple time point display of growing cell population segmented by color over time. Graph below tracks relative population numbers over time.
Image data courtesy of Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN.
Sequence Manager allows for variable time-lapse.
The Sequence Manager allows for time lapse of different durations, multiple nested loop cycles, and switching between galvanometer scanning and resonant scanning in the same experiment.
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 longterm 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.
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 MatTek.
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.
Users have their choice of two different types of scan units: galvanometer only with the FV3000 or galvanometer/resonant hybrid with the FV3000RS. The hybrid scan unit has galvanometer scanners for high-precision scanning, as well as a galvo/resonant scanner ideal for high-speed imaging. Galvanometer scanner enables Olympus super resolution technology (FV-OSR) yields resolutions down to 120 nm as well as high signal-to-noise, with precise tornado and multipoint stimulation and 100 ms switching time. Galvanometer scanning can achieve 16 frames per second at 2X zoom. The resonant scanner is capable of speeds ranging from 30 frames per second at 512 x 512 to 438 frames per second at 512 x 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.
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.
Click here for the details on FV3000 Hybrid Scan Unit
Loading the player… FV3000: Blood Flow Video |
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. |
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.
Optimized for Live Cell Imaging
Resonant scanning greatly reduces photobleaching and phototoxicity compared to standard galvanometer scans by preventing the excitation of fluorophores into triplet states that create reactive oxygen species. These features make live cell experiments more robust and reliable. The FV3000 Series has complete high and low range laser intensity control enabling the system to use the minimum required amount of laser power on samples. The optional Laser Power Monitor provides consistent laser power during long-term time-lapse imaging across multiple days.
The FV3000 Series employs Olympus’ TruSpectral detection concept. Based on patented* Volume Phase Hologram (VPH) transmission and an adjustable slit to control light, the spectral detection in FV3000 and FV3000RS is highly efficient, enabling users to select the detection wavelength of each individual channel to 1 nm.
* US8530824B/JP5541972B/EP2395380A
HSD makes it possible to view samples that were too dim to view with conventional equipment. The GaAsP PMT incorporates 2 channels with a maximum quantum efficiency of 45 %, and Peltier cooling reduces background noise by 20 % for high S/N ratio images under exceptionally low excitation light.
The FV3000 Series brings new levels of total system transmission efficiency, enabling every system to be completely spectral, improving overall sensitivity, and improving the signal-to-noise ratio for improved multi-color confocal imaging.
TruSpectral’s efficient design and software enable spectral detectors to run in multichannel mode for both live and postprocessing spectral unmixing with a multichannel lambda mode. Multichannel mode facilitates constant spectral unmixing during live cell experiments, separating complex fluorescence during acquisition. With up to 4 different dynamic ranges from the 4 different channels of array, even bright and dim spectral signals can be separated by adjusting the sensitivity of each detector independently.
Click here for the details on FV3000 High Sensitivity-Spectral Detector
Click here for the details on FV3000 Spectral Detector
Customizable and saveable layouts make it easy to tailor the interface to your workflow and experiment needs, from basic to complex.
Layout
Start by selecting your preferred display with specific tools for basic to complex acquisition.
Acquisition Condition
Reload settings that were ideal for your last experiment to provide consistency.
Acquisition
Activate basic to complex acquisitions with live ratio, intensity modulated display, quantitative region of interest (ROI) graphing or spectral unmixing display, and data backup for added security.
Viewer
Review data as it is generated. Generate 3D and 4D views and animations to explore and share data in depth.
Analysis
Extract data from images using online or offline processing. Analytical tools include Olympus super resolution technology (FV-OSR) and powerful cellSens software with features such as deconvolution, filtering, count and measure, and one-click macros.
The power of TruSpectral detection plus multichannel mode means live spectral unmixing can be performed during image acquisition, providing real-time processing of complex overlapping spectra.
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Live image before 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. |
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Live blind unmixed image.
Image data courtesy of Dr. Kazuhiro Aoki, Dr. Michiyuki Matsuda, Graduate School of Medicine, Kyoto University. |
Complex protocols are handled with ease, and real-time control helps provide microsecond accuracy of scans with millisecond accuracy over days of time-lapse.
Microplate imaging is important for many applications, and the Well Navigator provides sophisticated, intuitive controls for a wide range of cell culture vessels and custom plates. Multi-area timelapse and stitching provide robust and accurate time-lapse data.
Click here for the details on FV3000 Multi Area Time Lapse Software Module
The microscope comes equipped with a hard-disk drive (HDD) recording function. The images captured are stored automatically in the HDD. Large volumes of data, such as those obtained from long-term time-lapse imaging can be stored.
Images alone are not enough; with integrated cellSens Count and Measure analysis, the FV3000 Series can optimize images with deconvolution and analyze them with one-click macro functionality for a broad range of morphological measurements.
A spheroid image of a NMuMG cell line expressing Fucci2. Image data courtesy of Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN.
Click here for the details on cellSens Software
The FV3000RS includes an Intensity Modulated Display (IMD) function in the software that displays quantitative fluorescence ratio changes during both standard and high-speed acquisitions. This function is particularly useful for calcium and FRET imaging where a pure ratio display provides poor contrast in background areas.
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). Images courtesy of Shigeki Kiyonaka Ph,D, Yasuo Mori Ph,D Molecular Biology Field, Department of Synthetic Chemistry and Biological Chemistry, Kyoto University.
Cardiomyoctye Image data courtesy of Yusuke Niino and Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN.
High-speed scanning at low laser power to avoid phototoxicity often decreases the signal-to-noise ratio. With rolling average post-processing, users have the flexibility to adjust high-speed time-lapse images while maintaining time scale and keeping the original data.
(Left) Raw 30 fps data acquired at low laser power (0.05 %, 488 nm). (Right) Rolling average processing (10 frame) on 30 fps data acquired at low laser power.
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.
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 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.
Click here for the details on cellSens Software
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.
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.
Click here for the details on FV3000 Super Resolution Software Module
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.
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.
UPLSAPO30XS: For a broader view and greater depth
Magnification: 30X, NA: 1.05 (silicone oil immersion), W.D.: 0.8 mm,
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,
cover glass thickness: 0.13 - 0.19 mm, operating temperature: 23 - 37 °C
UPLSAPO60XS2: For 3D observations with superior resolution
Magnification: 60X, NA: 1.30 (silicone oil immersion), W.D.: 0.3 mm,
cover glass thickness: 0.15 - 0.19 mm, operating temperature: 23 - 37 °C
UPLSAPO100XS: For greater depth in closely defined regions
Magnification: 100X, NA: 1.35 (silicone oil immersion), W.D.: 0.2 mm,
cover glass thickness: 0.13 - 0.19 mm, operating temperature: 23 - 37 °C
In deep tissue observation, image quality depends on keeping the refractive index of the sample and immersion medium as close to each other as possible. When working with a silicone immersion objective, the difference between the refractive index of the samples and silicone oil is minimal. So it achieves brighter fluorescence images with higher resolution for deep tissue.
This oil immersion objective minimizes lateral and axial chromatic aberration in the 405 - 650 nm spectrum. Colocalization images are acquired reliably and images are measured with superior positional accuracy. The objective also compensates for chromatic aberration through near infrared up to 850 nm, making it the ideal choice for quantitative imaging.
Low Chromatic Aberration Objective
Magnification: 60X
NA: 1.4 (oil immersion)
W.D.: 0.12 mm
Chromatic aberration compensation range: 405 - 650 nm
Optical data provided for each objective.
Performance Comparison of PLAPON60XOSC2 and UPLSAPO60XO
A Z-drive guide installed near the revolving nosepiece combines high thermal rigidity with the stability of a wraparound structure to significantly reduce the impact of heat and vibration and improve the quality of time-lapse imaging.
Click here for the details on IX83 Inverted Microscope
The umbra unit is designed specifically for fluorescence observation. It efficiently blocks out room light, enhances the contrast of fluorescence, and enables clear fluorescence observation under bright conditions.
Hybrid Scan Unit (Resonant/Galvanometer)
The hybrid scanner combines the capabilities of a galvanometer scanner with a resonant scanner for high-speed imaging in the full field of view at 30 fps and up to 438 fps at 512 x 32. The Sequence Manager makes it simple to automatically switch between resonant and galvanometer imaging in the same experiment.
Click here for the details on FV3000 Hybrid Scan Unit
Galvo Scan Unit
The galvanometer-only scanner provides precision scanning from 1 fps at 512 x 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.
Click here for the details on FV3000 Galvo Scan Unit
High Sensitivity Spectral Detector (GaAsP PMT) with TruSpectral Technology
The 2-channel High Sensitivity Spectral Detector (HSD) employs the same Volume 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 2-channel spectral detector (SD) for a flexible dynamic range or a second 2-channel HSD unit for powerful 4-channel sensitivity.
Click here for the details on FV3000 High Sensitivity-Spectral Detector
Spectral Detector (Multialkali PMT) with TruSpectral Technology
The 2-channel SD employs efficient VPH transmission and an adjustable slit with 1 - 100 nm bandwidth from 400 - 800 nm detection. The multialkalai PMTs provide a broad dynamic range for detection up to 800 nm.
Click here for the details on FV3000 Spectral Detector
Main Laser Combiner
The main laser combiner is the heart of the laser system. Four standard lasers with an option to add a fifth laser or leave an open port to add an additional three diode lasers via the Sub Combiner.
Sub Laser Combiner
Add this optional combiner at any time with up to 3 diode lasers for a maximum of 7 laser lines in combination with the main laser combiner.
Click here for the details on FV3000 Laser Combiner
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 the microscope.
Light Source/U-HGLGPS
The pre-centered fluorescence illumination source requires no adjustment and has an average lifespan of 2,000 hours.
Click here for the details on U-HGLGPS
Transmitted Detector/FV31-LETD
This unit combines an external transmitted light 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.
Click here for the details on FV3000 Transmitted Detector
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.
Z Drift Compensator/IX3-ZDC2
The IX3-ZDC2 uses minimally-phototoxic IR light (laser class 1) to identify the location of the sample plane. One-shot autofocus (AF) mode allows several focus positions to be set as desired for deeper samples, enabling efficient Z-stack acquisitions in multi position experiments. Continuous AF mode keeps the desired 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. Furthermore, increased optical offset enables continuous AF over plastic vessels or with dry objectives. The IX3-ZDC2 is also compatible with silicone objectives (in AF mode).
Click here for the details on IX3-ZDC2
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.
IO interface Box/FV30-ANALOG
This unit supports electro-physiological experiments through analog inputs and TTL I/O support. The interface box converts voltage to images that can be treated in the same manner as fluorescence images.
Click here for the details on FV3000 IO Interface Box
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.
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.
| FV3000 | FV3000RS | ||
| Main Laser Combiner | 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, fiber 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
(Normal Imaging) |
Scanning Resolution: 64 x 64 to 4096 x 4096 pixels
Scanning Speed (One Way): 512 x 512 with 1.1 s - 264 s. pixel time: 2 µs - 1000 µs Scanning Speed (Round Trip): 512 x 512 with 63 ms - 250 ms, 256 x 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
(High-Speed Imaging) | - |
Scanning Resolution: 512 x 32 to 512 x 512 pixels
Scanning Speed: 30 fps at 512 x 512, 438 fps at 512 x 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 Unit for Scanner | Laser Power monitor, optional laser port | ||
|
High Sensitivity-
Spectral Detector | Detector Module | Cooled GaAsP photomultiplier, 2 channels | |
| 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) | ||
| Spectral Detector | Detector Module | Multi-Alkali photomultiplier, 2 channels | |
| 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 x 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 | ||
| 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. | |
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| 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) | |
Many biological processes can only be captured by video rate or higher imaging. The FV3000RS resonant scanner allows for imaging of highly dynamic processes such as calcium ion fluctuations, rapid muscle contractions, vesicle tracking and blood flow. In addition, it has been found that high speed scanning decreases photobleaching and phototoxicity by reducing the illumination time per area (photon flux) below the triplet dark state relaxation time, allowing for extended observation of live cells and tissues. The FV3000RS resonant scanner maintains a full 1X field of view at 30 fps and field number 18, providing greater context for rapid biological processes.
Loading the player… FV3000: Cardiomyocyte | FRET spectral look up table display of cardiac myocyte, CFP spectral look up table display of cardiac myocyte, Differential Interference Contrast (DIC) image of cardiac myocyte, Overlay, IMD ratio images of spontaneous Ca2+ oscillation in a beating rat cardiomyocyte expressing yellow cameleon. |
Loading the player… FV3000: Vesicle tracking |
Rapid movement of endosomes observed in the Drosophila embryo.
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Loading the player… FV3000: Blood Flow Video | 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. |
Loading the player… FV3000: Cell division resonant | 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. |
Loading the player… FV3000: 6x6 live cell resonant | Mouse Embryonic Fibroblast labeled with Silicon Rhodamine Docetaxol (Tubulin). Imaged with a 100X silicone objective, resonant scanning overview tiling overview. XYZT 6 x 6 areas. Courtesy Dr.Markus Delling, Harvard University. |
Loading the player… FV3000: Daphnia 10x DIC | Daphnea imaged in transmission detection, 30 fps. |
Multichannel TruSpectral Detection with 16-Channel Unmixing
TruSpectral’s efficient design and software enable spectral detectors to run in array mode for both live and post-processing spectral unmixing with a multichannel lambda mode. Multichannel mode facilitates constant spectral unmixing during live cell experiments, separating complex fluorescence during acquisition. With up to 4 different dynamic ranges from the 4 different channels of array, even bright and dim spectral signals can be separated by adjusting sensitivity of each detector independently.
Loading gallery... Loading...  | 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. |
Loading gallery... Loading...  | Live image before 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. |
Loading gallery... Loading...  | Live blind unmixed image. Image data courtesy of Dr. Kazuhiro Aoki, Dr. Michiyuki Matsuda, Graduate School of Medicine, Kyoto University. |
Loading gallery... Loading...  | Spectrally deconvolved image of 9 channel lambda stack. |
Loading the player… FV3000: Multi Lambda | Raw spectral acquisition. |
Traditional confocal systems focused solely on micro imaging and neglected low magnification capabilities, however increasingly biologists have come to need the ability to place microscopic observations in the context of larger tissues and whole organisms. The FV3000 has been designed to enable macro scale imaging from 1.25X, while retaining the high precision capabilities up to super-resolution imaging. Imaging with low magnification speeds up workflow for broad overviews of samples, and eliminates stitching artifacts that often occur from sample mounting tilt and other factors.
Loading gallery... Loading...  | Mouse kidney prepared by Dr. Mike Davidson. Images presented with lasting gratitude for his lifetime commitment to science and microscopy. |
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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. |
Loading the player… FV3000: Whole rat embryo | 2 x 2 tiled image of whole rat embryo, 20 mm total field of view. H&E fluorescence with 640 nm laser diode. |
Multi-dimensional Time-lapse Imaging with Outstanding Positional Accuracy
The FLUOVIEW FV3000 can be used for ideal multi dimensional time-lapse imaging during confocal observation, using multi-area time-lapse software to control the motorized XY stage and IX3-ZDC2 Z-drift compensator.
Significantly Improved Multi-Point Time-Lapse Throughput
Equipped with motorized XY stage for repeated image acquisition from multiple points scattered across a wide area. The system efficiently analyzes changes over time of cells in several different areas capturing, large amounts of data during a single experiment to increase the efficiency of experiments. Microplates can be used to run parallel experiments, which significantly improves throughput for experiments that require long-term observation.
Supports repeated image acquisition from multiple areas in a single microplate well.
Loading gallery... Loading...  | Loading gallery... Loading...  | Loading the player… FV3000: Using Methods Similar to Random Access Scanning |
Pancreas vasculature visualized after PFA fixation and SCALEVIEW-A2 clearing treatment
Loading the player… FV3000: The mouse pancreas which cleared by SCALEVIEW-A2 | Blood vessels labeled with Texas Red-labeled lectin via transcardial perfusion. Imaged with UPLSAPO30XS objective. Image size 425 um x 436 um (616 x 633 pixels) Z: 438 um (219 slices). Laser: 559 nm.
Image data courtesy of:
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High-level Magnification With High Resolution for The Broad-scope Imaging of Large-scale Specimens
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)
Automated from 3D Image Acquisition to Mosaic Imaging
Multi-area time-lapse software automates the process from 3D image acquisition (using the motorized XY stage) to stitching. The software can be used to easily register wide areas, and the thumbnail display provides a view of the entire image acquired during the mosaic imaging process.
Multi-point Imaging and Montage Imaging
Tiling image acquisition by equipping the system with a motorized XY stage, repeated image acquisition of multiple points located over a large specimen can be automated. Image made by acquiring images at multiple points of a mouse brain section (YFP) and integrated.
Selectable Rendering Modes
Interactive Volume Rendering
Using the interactive volume rendering method with the 3D display function, the angle of a 3D rendered image can be freely changed to the direction you wish to see by moving the mouse. A variety of display functions are available, including extended focus images and the ability to display a cross section at an arbitrary location.
4D Animation Creation Function
Loading the player… FV3000: 4D Animation Creation Function | 3D structures which change with time can be animated for images acquired with XYZT. |
Brightness Compensation Function for the depth direction
When acquiring 3D images, the images become dimmer with depth. The system is equipped with a function for increasing laser intensity or PMT sensitivity with depth so that images of deeper regions can be acquired with equal brightness.
Loading gallery... Loading...  | A wide range of live and post processing analysis functions are available to enhance data production:
Highlighted cellSens Image Processing and Analysis (Please click link to see extensive list of cellSens functions)
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Click here for the details on cellSens Software
A wide range of scanning modes are available for resonant and galvanometer scanning. Please see Specification Tab for additional details.
Resonant Scanning
Resonant scanning allows for high speed acquisitions, improving dynamic measurements and reducing photobleaching and phototoxicity in a wide field of view. With full 1 x field scanning at 30 frames per second, clip scanning at up to 438 frames per second, or line scanning at up to 7.87 kHz, the FV3000RS resonant scanner provides blazing speed for dynamic imaging and analysis.
Galvanometer Scanning
Galvanometer scanning provides high precision and signal to noise, and an increased range of scan paths and stimulation capabilities. The FV3000 series galvanometer scanner also sports an Interlace scan mode capable of 16 frames per second with only a 2X zoom factor.
Live Graph Function
Changes in fluorescent intensity in a region designated as ROI are plotted in real-time during image acquisition.
Live Tile Mode, Look Back Function
This allows the user to check the acquired images during time-lapse experiment. Adjustment of focus and/or image brightness can be done during rest time.
Trigger Function
The system also has a trigger function for synchronizing scanning with external devices. Scanning can be started/stopped with a trigger signal and it is also possible to use an external trigger for each image frame acquisition.
Optional Laser Power Monitoring Function
Feedback is applied to the laser output so that the sample is always exposed to a fixed level of excitation light and the amount of fluorescence can be measured accurately.
Fast sequential photostimulation can be done using the galvanometer scanner with as little as 100 ms delay. Applications such as FRAP, FLIP and photoconversion in complex areas can be achieved in live samples.
Photoconversion (Kaede)
The Kaede protein is a typical photoconvertible protein, which is a specialized fluorescent protein that changes color when exposed to light of a specific wavelength. When the Kaede protein is exposed to laser light, its fluorescence changes from green to red. This phenomenon can be used to mark individual Kaede-expressing target cells among a group of cells, by exposing them to laser light.
This fluorescent protein is made through cloning from Trachyphyllia geoffroyi. It emits a strong green light after synthesis, but when stimulated by ultraviolet laser illumination, changes its color from green to red, like a maple tree in autumn. Its name is derived from this characteristic ("kaede" means "maple tree" in Japanese). When violet (or ultraviolet) laser illumination is directed onto a Kaede-expressing cell, diffusion of the reddish Kaede can be monitored throughout the whole area of the cell, providing an easy and accurate method to label the entire cell.
Kaede-expressing astroglia cells are stacked on the Kaede-expressing neurons. By illuminating two colonies with a 405 nm laser, the Kaede color can be photoconverted from green to red. The glial cells in contact with the neurons are observed while they are forming colonies and extending their processes, and the nuclei of these colonies can also be observed.
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