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Overview
Next Generation FLUOVIEW for the Next Revolutions in Science
The FLUOVIEW FV3000 series of confocal laser scanning microscopes meets some of the most difficult challenges in modern science. Featuring the high sensitivity and speed required for live cell imaging as well as deep tissue observation, the FV3000 confocal microscope enables a wide range of imaging modalities, including macro-to-micro imaging, super resolution microscopy, and quantitative data analysis. Choose between upright and inverted microscope frames suited for a range of life science applications, including developmental biology, stem cell research, electrophysiology, cancer research, slide imaging, and more.
TruSpectral High-Sensitivity Multichannel Imaging
Using proprietary spectral detection technology, the FV3000 confocal microscope's TruSpectral detectors combine high sensitivity with spectral flexibility to detect even the dimmest fluorophores.
- Up to 3X more light transmission vs. traditional spectral detection technology
- Independently adjustable channels to optimize signal detection for each individual fluorophore
- Lambda scanning mode enables accurate spectral unmixing of complex overlapping fluorescent signals
- Variable barrier filter mode provides simultaneous four-channel image acquisition, with up to sixteen channels in virtual channel mode
Macro-to-Micro Imaging and Super Resolution Microscopy
The FV3000 microscope’s macro-to-micro workflow provides a roadmap for data acquisition, enabling you to see data in context and easily locate regions of interest for higher resolution imaging.
- Use a low-magnification 1.25X or 2X objective to quickly capture a large field of view (FOV) map of whole specimens
- Identify regions of interest on the Overlay Map, then switch to a higher magnification objective for high-resolution confocal imaging down to 120 nm with Olympus Super Resolution technology (FV-OSR)
- Finalize your acquisition and get publication-ready microscope images with TruSight image processing
Mouse brain hemisection embedded for expansion microscopy (pre-expansion), labeled with secondary antibodies against GFP (Alexa Fluor 488, green), 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 constrained 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.
Hybrid Scanning for High-Speed Imaging and Increased Productivity
The FV3000 Hybrid Scanner provides two scanners in one for enhanced confocal imaging capabilities.
- The FV3000RS hybrid scan unit uses a galvanometer scanner for precision scanning as well as a resonant scanner, ideal for high-speed imaging of live physiological events
- Capture video-rate images with a large FOV using the resonant scanner, featuring speeds from 30 frames per second (fps) at FN 18 all the way up to 438 fps using clip scanning
- Use the resonant scanner to observe fast phenomena, such as a beating heart, blood flow, or calcium ion (Ca2+) dynamics inside cells
- Switch between the galvanometer scanner and resonant scanner with the click of a button
FV3000: Cardiomiocito
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FV3000: Dudi-158V-XYT-0001_00000
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Accurate Time-Lapse Imaging
Time-lapse experiments require consistent focus and low phototoxicity to the sample.
- Olympus’ TruFocus unit helps maintain focus during live cell imaging despite changes in temperature or added reagents
- The FV3000 microscope’s high-sensitivity detector requires significantly less laser power while the resonant scanner reduces laser illumination time, lowering phototoxicity for more physiologically accurate confocal imaging data
Deep Tissue Observation with Silicone Objectives
The refractive index of silicone oil is close to that of living tissue, enabling high-resolution observation deep inside tissue with minimal spherical aberration.
- Refractive index match delivers an ideal focal volume, resulting in perfect volume reconstruction and enabling high-resolution confocal imaging of large living organisms
- Long working distances enable detailed microscopic imaging at depth
- See data unfold in real-time and easily observe structures with 3D reconstruction software
3D image of chick ciliary ganglion cleared by a tissue clearing reagent.
Image data courtesy of Dr. Ryo Egawa. Tohoku University Graduate School of Life Science.
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Applied Technologies
TruSpectral Detection
Groundbreaking TruSpectral detection technology offers outstanding results compared to previous generations of spectral detection units. Featured on every channel in the FV3000 microscope, TruSpectral detection technology combines the flexibility of a spectral detector with the sensitivity of a filter-based detector.
How TruSpectral Detection Technology Works
Based on patented volume phase hologram (VPH) transmission for high efficiency, TruSpectal detection uses an adjustable slit to select the detection wavelengths of each individual channel to 2 nm.
Sensitivity and Accuracy
Integrated in all FV3000 confocal microscopes, TruSpectral detection technology enables much higher light throughput compared to conventional spectral detection units. The volume phase hologram diffracts light with up to three-fold higher transmission efficiency vs. reflection gratings. The result is excellent multicolor fluorescence microscopy images of live and fixed tissue.
Enhanced Quantum Efficiency
The FV3000 microscope's high-sensitivity detector (HSD) enables you to view samples whose emission is too weak to view with conventional detectors. The HSD unit incorporates two GaAsP channels with a maximum quantum efficiency of 45% and Peltier cooling that reduces background noise by 20% for high signal-to-noise (S/N) ratio images under very low excitation light. HSD units can be combined to enable four-channel GaAsP imaging with the FV3000 system.
Original
Sensitivity Adjustment of Each Channel
Spectral Unmixing
Multichannel TruSpectral Detection with Sixteen-Channel Unmixing
TruSpectral detection works independently on all microscope channels, enabling true simultaneous multichannel lambda scanning on up to four channels. Multichannel lambda mode facilitates both live and post-processing spectral unmixing for excellent spectral separation results. With up to four dynamic ranges, bright and dim signals can be optimally separated by independently adjusting the sensitivity of each detector.
Spectral Unmixing
The FV3000 system’s spectral deconvolution algorithm enables overlapping spectra to be separated based on the spectral information from lambda stack images. Fluorescence cross-talk between channels can be eliminated by the unmixing algorithm during both live image acquisition and post-processing, enabling clean separation of up to sixteen fluorophores.
Spectral unmixing of a PtK2 cell labeled with YOYO-1, Alexa Fluor 488, Rhodamine-phalloidin, and MitoTracker Red using multi-channel lambda stack image.
Platelets bound to a thrombosis in the blood vessel of a mouse. Images taken at 30 fps in full frame by a resonant scanner with 2 CH GaAsP PMTs.
Image data courtesy of: Dr. Takuya Hiratsuka, Dr. Michiyuki Matsuda, Graduate School of Biostudies, Kyoto University.
Two Scanner Options
Choose between two scan units: a traditional galvanometer scanner (FV3000) or a hybrid galvanometer/resonant scanner (FV3000RS).
- Using the galvanometer scanner and Olympus super resolution technology (FV-OSR), obtain resolutions down to 120 nm with a high signal-to-noise ratio
- Galvanometer unit offers flexible scanning options, including precise tornado scanning, as well as multipoint stimulation with 100 ms switching time
- Hybrid scan unit features both a galvanometer scanner for high-precision scanning as well as a resonant scanner for high-speed imaging
- Resonant scanner enables you to capture 30 frames per second with a full field of view at 512 × 512 pixels, or up to 438 frames per second by clipping down in Y to capture critical live physiological events, such as calcium ion flux
No Compromise Between Speed and Field of View
Many high-speed scanning methods restrict the field of view, limiting your ability to examine large areas with multiple cells. The resonant scanner in the FV3000RS microscope maintains a full 1X field of view with FN 18, even at a video rate of 30 frames per second. Clipping the Y axis enables speeds up to 438 frames per second.
Rolling Average Processing
While high-speed scanning with low laser power minimizes phototoxicity, it often decreases the signal-to-noise ratio, making it difficult to obtain high-resolution time-lapse images. With rolling average processing, you can adjust high-speed time-lapse images to achieve better signal-to-noise ratios while maintaining the time scale and keeping the original data.
(Right) Rolling average processing (10 frames) on 30 fps data acquired at low laser power.
Macro-to-Micro Observation
See data in context with macro-to-micro observation. With the FV3000 system’s redesigned light path, generate detailed overview images with magnifications as low as 1.25X, then easily identify structures to image at higher magnification. Mosaic imaging enables you to acquire continuous 3D (XYZ) and 4D (XYZT) images of adjacent fields of view. The entire process from image acquisition to stitching can be fully automated, saving time and generating more meaningful data.
TruSight Deconvolution: Image Processing for Higher Resolution
Remove blur and obtain clearer, sharper images with TruSight deconvolution. Specialized cellSens algorithms for the FV3000 confocal microscope enable a seamless workflow from acquisition to publication with the click of a button. Take advantage of GPU processing for even faster results.
Image of a GATTA-SIM nanoruler
Left: Raw confocal image / Right: Image with TruSight
0.5 AU Confocal Image
0.5 AU Confocal Image Deconvolved with cellSens Advanced Deconvolution
Olympus Super Resolution Plus cellSens Advanced Deconvolution. Note clear separation of punctate stains with OSR.
Olympus Super Resolution (FV-OSR) Technology with Up to Four Simultaneous Channels
Suitable for colocalization analysis, the Olympus Super Resolution imaging module can acquire four fluorescent signals either sequentially or simultaneously with a resolution of approximately 120 nm, nearly doubling the resolution of typical confocal microscopy.
- Requires no special fluorophores and works with a wide range of samples
- Easy to use with minimal training
- Add FV-OSR to any FV3000 confocal microscope system to achieve super resolution microscopy
Learn more about the FV3000 Super Resolution Software Module
TruFocus Time-Lapse Imaging
TruFocus technology makes multi-position experiments and long-term time-lapse imaging more robust and reliable. TruFocus technology uses a minimally phototoxic infrared laser (Class 1) to identify the location of the sample plane and offers two modes for maintaining focus.
- Single shot autofocus (AF) mode allows you to set several focus positions, enabling efficient Z-stack acquisitions in multi-position experiments
- Continuous AF mode operates continually to keep the desired plane of observation precisely in focus, making it ideal for time-lapse measurements by avoiding focus drift due to temperature changes or added reagents
- TruFocus is compatible with a wide range of confocal imaging conditions, including dry objectives, plastic vessels, and silicone objectives
TruFocus: Z-Drift Compensator
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High-Performance Microscopic Imaging with X Line Objectives
Providing broad chromatic aberration correction, uniform images, and a high numerical aperture, X Line objectives enhance the FV3000 confocal microscope’s image quality.
Deep Tissue Observation with Silicone Immersion Objectives
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
- Silicone oil does not dry out or harden, so you don’t need to refill oil during extended time-lapse observations
Silicone Oil Immersion Objectives: For Live Cell Imaging
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PLAPON60XOSC2
Performance Comparison of PLAPON60XOSC2 and UPLXAPO60XO
Enhance the Reliability of Colocalization Analysis with a Low Chromatic Aberration Objective (PLAPON60XOSC2)
This oil immersion objective minimizes lateral and axial chromatic aberration in the 405–650 nm spectrum, enabling you to acquire reliable colocalization images and measure objects with positional accuracy. The objective also compensates for chromatic aberration through near infrared (up to 850 nm), making it ideal for quantitative microscopy imaging.
- Magnification: 60X
- NA: 1.4 (oil immersion)
- WD: 0.12 mm
- Chromatic aberration compensation range: 405–650 nm
- Optical data provided for each objective
Optimized Configurations for Electrophysiological Experiments
Synchronize confocal imaging events with electrophysiology equipment via the trigger signal I/O interface box. The I/O interface box also converts voltage signals into images that can be treated in the same manner as fluorescence images. This enables you to capture voltage-based images in synchrony with photostimulation by the confocal scanner.
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Software
Intuitive Software
The FV3000 software streamlines your entire confocal imaging workflow, from acquisition to analysis. Customizable and savable layouts make it easy to tailor the interface to your workflow and experiment needs, no matter the level of complexity.
Fucci induced spheroid of the HT29 cell line
Image data courtesy of: Yuji Mishima, Ph.D., Kiyohiko Hatake M.D., Ph.D. Clinical Chemotherapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research.
Live 3D Rendering
See your data unfold in real time with the FV3000 software’s live 3D image display function. 3D images can be constructed during image acquisition and shown as live images.
Multi-Area Time-Lapse and Microplate Imaging
The Multi-Area Time-Lapse (MATL) module provides robust and accurate time-lapse data through precise control over motorized stage movements, enabling you to generate detailed overview microscope images to see data in context. Pair the MATL module with the well navigator module to achieve more functionality with sophisticated, intuitive controls for a wide range of cell culture vessels and custom microplates.
Reduce Complexity with the Sequence Manager
The Sequence Manager software module handles complex protocols with ease. Multiday time-lapse experiments are controlled with microsecond scan accuracy and millisecond sequence execution accuracy. Various complex protocols can be performed, including:
- Switching between high and low magnification
- Variable-interval time-lapse experiments
- Photostimulation between imaging by FRAP or FRET (acceptor photobleaching)
Quantitative Microscopy Image Analysis
The FV3000 confocal microscope incorporates a suite of optional analysis functions to complete your imaging workflow and provide quantitative data.
- Count and Measure: calculate the number, size, luminosity, and morphology of cell segments
- Colocalization: analyze overlapping fluorescent spectra
Count and Measure
Sample courtesy of: Hidetaka Kosako, Fujii Memorial Institute of Medical Sciences, Tokushima University
FRET and FRAP Experiments
The FV3000 microscope paired with the cellSens Life Science Analysis module enables easy acquisition and analysis of FRET and FRAP experiments.
- FRAP: estimate τ/2 and the mobile/immobile fraction by fitting the curve of luminosity change caused by fluorescence recovery after bleaching
- FRET: measure FRET efficiency by acceptor photobleaching, ratio imaging, and sensitized emission
Example of FRET analysis (acceptor photobleaching)
Example of FRAP analysis
Ratio Imaging and Intensity Modulated Display (IMD)
The FV3000 microscope's ratio imaging analysis software includes an Intensity Modulated Display (IMD) function 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.
Before CCCP treatment
After CCCP treatment
tsGFP1-mito reveals heterogeneity in mitochondrial thermogenesis in HeLa cells.
The images of the ratio (ex 405 nm/ex 488 nm) in tsGFP1-mito-expressing cells before and after CCCP treatment at 37 °C (98.6 °F). 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.
(Left) CFP, (Right) YFP FRET
(Left) Raw CFP/YFP ratio, (Right) 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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Bioluminescence of RA-induced differentiating cells at day 12 from Bmal1:luc stably transfected ES cells
Image data courtesy of: Kazuhiro Yagita, M.D. Ph.D. Department of Physiology and Systems Bioscience, Kyoto Prefectural University of Medicine
Reference: Proc Natl Acad Sci U S A. 107(8): 3846–3851 (2010)
Object Tracking
Automatically detect, track, and analyze moving objects in time-lapse images using the cellSens Object Tracking module. The tracking function provides a powerful and intuitive tool to quantify dynamic processes, such as cell movement and division.
Remote Development Kit (RDK) Module
The Remote Development Kit enables remote control and programming of certain FV3000 microscope functions in languages like Python, C++, and Matlab. The RDK module is a powerful tool for users with programming experience to unlock even greater potential behind the system.
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Configurations
Choose the Configuration that Suits Your Application
Inverted microscope
- Suitable for observing cells cultured in a vessel
- TruFocus unit enables time-lapse observations with consistent focus
- Add a stage-top or full enclosure incubator to maintain the environmental conditions of cultured cells
Upright microscope (configured for slide imaging)
- Optimized for fixed tissue and glass slide specimens
- Motorized 7-position nosepiece and condenser enable automated transitions from low to high magnification for macro-to-micro applications
Upright microscope (configured for electrophysiology)
- Ample space around the objectives enables easy installation of patch-clamp devices
- Add extra space by lowering the stage position for experiments that require large sample handling
- Swing and slider nosepieces are available so objectives can be easily changed without interfering with the patch-clamp setup
Componentes
- 2 channel high-sensitivity Olympus-patented TruSpectral detection
- Cooled GaAsP PMT
- New high-throughput TruSpectral detection system (Multi channel lambda scanning)
- Variable Barrier Filter (VBF)
FV3000 High Sensitivity-Spectral Detector
- Simple hardware interface
- External data processing via two additional channels
- Easily output scan timings and triggering
- I/O signal processing
- Modular compact solid-state laser system
- Stable laser power
- Ultra-fast and flexible laser control
- One broadband fiber for all laser lines
- Low operating cost
FV3000 Main Laser Combiner and Sub Laser Combiner
- 2 channel high-sensitivity Olympus-patented TruSpectral detection
- Multi-alkali PMT
- New high-throughput TruSpectral detection system (Multi channel lambda scanning)
- Variable Barrier Filter (VBF)
- Two-in-one module
- Multichannel image acquisition
Software
- Multi area time lapse
- 3D mosaic imaging and stitching
- Long-duration time lapse system without focal drift
FV3000 Multi Area Time Lapse Software Module
- Multipoint scans
- Mapping scans
- Create activity maps
FV3000 Multi Point and Mapping Software Module
- Programmable interface to FV31S-SW
- Excellent 120 nm optical resolution with FV-OSR
- Simultaneous multicolor super resolution
- Enhanced cell and tissue imaging
- High sensitivity and low noise with cooled detector technology
- 3D image recognition from whole structure to sub-cellular
- Statistical analysis
- Variety of default assays
Scanners
- Highly reflective silver-coated scanning mirrors
- Tornado scanning
- Optional laser power monitoring
- Motorized ND filter
- Fast high-precision scanning
- Highly reflective silver-coated scanning mirrors
- Tornado scanning
- Optional laser power monitoring
- Motorized ND filter
Especificaciones
| 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 the sub laser combiner or optional laser unit | |
|---|---|---|---|
| Optional Laser | Sub Laser Combiner |
Maximum 3 laser units as follows:
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 using an 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 × 64 to 4096 × 4096 pixels
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
(High-Speed Imaging) | - |
Scanning Resolution: 512 × 32 to 512 × 512 pixels
Scanning Speed: 30 fps at 512 × 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) | ||
| System Control | Control Unit |
OS: Windows® 7 Professional 64-bit (English version), Windows 10 Professional 64-bit ;
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 adaptor to the optical port of the scan unit, motorized switching between the LSM light path and fluorescence illumination | ||
| Transmitted Light Detector Unit | Module with integrated external transmitted light photomultiplier detector and LED lamp, motorized switching | ||
Microscope
| Inverted frame | Upright frame (for imaging) | Upright frame (for electrophysiology) | ||||||
| Microscope Frame |
Motorized inverted microscope
IX83 (IX83P2ZF) |
Motorized fixed stage upright microscope
BX63L |
Motorized fixed stage upright microscope
BX63L | |||||
| Revolving Nosepiece | Motorized sextuple revolving nosepiece | Motorized septuple revolving nosepiece |
Coded swing nosepiece
Coded slider nosepiece | |||||
| Condenser | Motorized long working distance condenser | Motorized universal condenser | Manual long working distance condenser | |||||
| Focus Stroke |
Built-in motorized nosepiece focus
Stroke: minimum increment: 0.01 μm | |||||||
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 | Region and line measurements, intensity plot over time/Z, colocalization analysis. | |
| Statistical Processing | 2D data histogram display. | |
| Optional Software | Motorized-stage control | |
Recursos
- Inicio
- Productos
- Microscopios de barrido láser
- Microscopio confocal de escaneo láser
