Multi Mode Multiphoton Scanning Microscope
With the Olympus FVMPE-RS, countless possibilities for deep tissue observation are finally realized. The system delivers unmatched high-speed imaging, essential for capturing the dynamic in vivo response, with fine laser control pinpointing the precise site for optimum excitation efficiency - even deep within the sample. Accompanied by high S/N-ratio imaging and dedicated Olympus multiphoton objectives, efficient detection of scattered fluorescence photons is also ensured. Optimized for reaching greater imaging depths, the system also features simultaneous dual-wavelength excitation (up to 1300 nm). Visible or multiphoton laser light stimulation and the ability to synchronize with patch clamp data are also possible. In essence, the Olympus FLUOVIEW FVMPE-RS unites high-speed, deep observation capability with multicolor imaging and powerful laser light stimulation, offering a no compromise solution.
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A High-Speed Scanner Providing Unique 438 fps at 512 × 32 Scan Performance
The scanner unit combines a newly developed high-speed resonant scanner with a conventional galvanometer scanner to provide high-speed and high-definition imaging in a single system. High-speed imaging delivers 30 fps at 512 - 512 at full field of view (FN 18), while clip scans optimize return time to achieve 438 fps at 512 × 32 pixels - making it possible to capture rapid calcium channel reactions and membrane potential-sensitive dyes in action.
A Proprietary Silver Coating Improves Excitation Efficiency by 50 %*
Silver-coated scanner mirrors achieve extremely high reflectance characteristics across a broad wavelength range from visible to near-infrared. Total reflectance for the XY scanner is enhanced by more than 25 % in the near-infrared range compared to conventional aluminum coating mirrors, and this increased reflectance provides more than a 50 % improvement when converted to multiphoton excitation efficiency. The result is a highly effective apparatus that delivers the superior power needed
for deep in vivo probing.
*Compared to standard aluminum coating.
A Cooled, High-Sensitivity GaAsP Detector Acquires High S/N Images
High S/N imaging can be acquired even under faint fluorescence through the use of gallium arsenide phosphide (GaAsP) in the photomultiplier tube (PMT) - delivering greater quantum efficiency than multialkali PMT along with Peltier cooling that improves S/N even further.
Laser with Negative Chirp Improves Excitation Efficiency at the Focal Plane
A laser beam with optimally adjusted pulse widths can be delivered to the focal plane, thanks to the application of negative dispersion that perfectly corresponds to the magnitude of the pulse-width dispersion generated during transmission through the microscope optics. The result is brighter images without needing to increase laser power, sample heating, photobleaching and phototoxicity.
Deep Focus Mode Elevates Light-Condensation Performance for Specimens with Heavy Scattering
A newly developed Deep Focus Mode responsively adjusts the laser beam diameter in accordance with laser scattering conditions across specimens. For in vivo specimens with heavy laser scattering, more excitation photons reach deep sites with the Deep Focus Mode and produces brighter high-resolution images.
Depth-Brightness Compensation Keeps Brightness Consistent from the Surface to Deep Levels
When observing thick specimens, images can often get darker as the focal point goes deeper. But with depth-brightness compensation, detector sensitivity and laser power are constantly retuned to keep brightness at a consistent level.
Optics with Outperforming New IR Coating for Applications from 400 nm to 1600 nm
An innovative IR coating (1600 coating) for the 25× dedicated multiphoton objective range and scanner optics further refines deep observation quality. This coating with improved long wavelength transmittance enables excitation without a decrease in laser power, even at deep sites. Since transmittance at 405 nm also remains high, the feature is suited to uncaging applications that employ a 405 nm laser.
Deep Observation of In Vivo and Fixed Transparent Specimens Through Dedicated Multiphoton Objectives with a Maximum Depth of 8 mm
The 25x (XLPLN25XWMP2) water immersion objective with a working distance of 2 mm delivers a high resolution and a wide field of view for the deep observation of live specimens. Three other objectives in the same family with working distances of 4 mm and 8 mm deliver maximum performance with fixed transparent specimens for high-definition observation at deep levels and accommodating various immersion solutions. A water to oil multi-immersion 10x objective is added to the growing Olympus optics list with a large field view and 8 mm of working distance. All of these objectives feature correction collars that allow them to correct spherical aberration generated by the difference in refractive index between the immersion solution and the specimen - forming optimal light-condensed spots without energy density loss, even during observations deep within the specimen. Furthermore, each objective features a wide field design that permits the efficient acquisition of scattered fluorescence photons for bright observations.
Silicone Immersion Objectives for Live Imaging
This immersion objective is designed exclusively for use with silicone oil, which has a refractive index even closer to live cells than that of water. The objective features a large numerical aperture and wide-ranging transmission capability from UV to IR for use in both multi photon and single photon microscopy. Time lapse observations become more reliable and less elaborate, because silicone oil does not dry at 37ºC and its refractive index remains constant. This objective also offers a long working distance to enable observation at deeper tissue levels and across wide field of views. In a nutshell, this silicone objective offers a comprehensive solution for both macro- and deep-tissue observation in the fields of developemental and regenerative science.
Multi-wavelength Excitation And Multiphoton Imaging with Precision Imaging Co-registration
Multichannel, multiphoton excitation imaging is accomplished with a dual-wavelength IR pulsed laser or two independent IR pulsed lasers - enabling simultaneous excitation of chromophores with different wavelengths. Thanks to the flexible and precise IR introduction optics both lines are accurately merged. Simultaneous excitation provides perfected registration and balanced images for different chromophores.
InSight DeepSee And Combination with Other Lasers Supports Simultaneous Two Laser Line Excitation and Extended NIR Multiphoton Imaging
The InSight DeepSee pulsed IR laser systems ideally support multiphoton imaging with excitation from 680 - 1300 nm. The Dual-line version of the InSight DeepSee system offers two laser beam outputs: the main output with a tunable line from 680 - 1300 nm and a second output at 1040 nm. Higher laser power beyond 1000 nm provides a host of new multiphoton imaging capabilities, covering a variety of dyes and fluorescence proteins and Third Harmonic Generation imaging without UV damage.
Autocorrect for Laser Beam Misalignment and Pixel Shift with Quadralign 4 Axis Auto Alignment
Multicolor multiphoton laser acquisition provides optimized excitation of different fluorophores, reducing channel crosstalk and photobleaching in the single beam excitation method due to the need to choose a suboptimal middle wavelength for excitation. To ensure proper colocalization of fluorescent signals, the Quadralign 4 axis auto alignment is incorporated into 2 horizontal and 2 angular axes per laser line, and single-click compensation is also enabled for laser beam position as well as incident laser angle - a common cause of pixel shift by tuning wavelength. Saving time and effort, this auto alignment mechanism tunes the optical axes of the lasers to the laser wavelength used during multicolor excitation. Software-based fine-tuning is also available.
Software Architecture Supports Massive Data Needs
Smooth, 3D-rendered display is possible for massive Z-Stack data comprising high definition images captured from the sample surface to deep sites. Key frame registration is also available, making it easy to create animated views of 3D images that zoom and transition to different camera angles.
Tiling Significantly Extends the Imaging Range
The tiling function scans multiple adjacent views and stitches them together to build a large image beyond the physical field of view. Use of a motorized stage supports tiling for an even wider field of view, while the mapping feature makes it easy to locate a specific cellular position within the resultant large image.
Software Sequencer Control Allows Complex Experimental Sequence with Precision Timing over Days
Microsecond repeatability precision provides the power needed for precise control of triggering and point of stimulation. The optional sequencer manager enables extra long-term (two-week) procedures for complicated observational testing that requires switching between different imaging tasks. Even in extra-extended lab work cycles, repeatability retains millisecond precision. Microsecond repeatability precision is critical for many applications requiring high speed.
This is particularly true for electrophysiology and optogenetic stimulation, where microsecond timing can mean the difference between observing synchronous and asynchronous stimulus responses.
For extra-long (two-week) acquisitions with complicated experimental procedures that require switching between different imaging tasks, the optional sequencer manager can still maintain millisecond precision, ensuring data integrity in the most demanding in vivo and in vitro experiments.
Light-Stimulation SIM Scanner from the Visible to IR Range
A laser light stimulation scanner can be installed separately - enabling optogenetics laser light stimulation of channel rhodopsin (ChR2) and halorhodopsin (NpHR) with simultaneous real-time imaging of neural cell activity with a visible or IR-range laser.
Wide Choice of Scan Modes
Analog Unit Synchronizes Electro-Physiological Data and Laser Light Stimulation
Electro-physiological experiments are enabled through analog inputs and TTL I/O support. The analog unit converts voltage to images that can be treated in the same manner as fluorescent images - enabling multiple channel light-stimulated electrical signals measured with patch clamps to be synchronized with image capture and displayed as a pseudo-color intensity overlay.
3D Multi Point Measurement Provide Rapid Intensity Measurement with High Signal to Noise
Raster scanning takes time. With MMASW(Multi-Point Mapping Software) multi-point acquisitions you can eliminate that time by placing the laser only where needed, retaining high signal to noise output and allowing you the freedom to choose and optimize your scan path. A scan of multiple positions can go as fast as 101 Hz, and each position can gather signals at up to 50,000 Hz, providing you with highly relevant physiological data.
MMASW is designed for extremely fast functional measurements in living cells or tissues where researchers use light to probe fluorescent intensity changes. Resonance scanners or acousto-optic deflectors (AODs) are intrinsically less sensitive due to their very short integration times and the number of photons that can be detected. By retaining high integration time per position where it is needed in a single point laser scan, the multi-point mode allows greater multiphoton depth penetration
and signal-to-noise. Each point can also be expanded to an array for larger area stimulation or detection.
Synchronize your measurement scans with simultaneous stimulation using the SIM scanner and free your functional imaging.
3D Mapping Stimulation Creates Reaction Maps Based on Multiple Coordinates
Highly targeted laser light stimulation is achieved through dividing the observation domain into a grid and laser irradiating each specific area in a software-controlled sequence while eliminating adjacent areas from stimulation. The Z position setting is available to enable stimulation at a depth different from that of the imaging layer. Changes in intensity during stimulation can also be mapped to the image and reaction maps can be created for multiple coordinates.
This Upright Frame is completely dedicated to multiphoton microscopy. Providing space for large samples, a high degree of motorization and nosepiece focus control enables the stage and your sample to remain fixed and stable.
With its ultra-stable arch-like structure, the new Gantry Frame offers tremendous space beneath the objective lens along with a high degree of flexibility to suit different samples. This is ideal for in vivo observation requiring maximum space.
The Inverted Frame is ideal for the time lapse observation of thick, living specimens such as tissue cultures, and three-dimensional cell cultures.as The inverted frame system also finds utility in intravital time lapse observation of organs and tissues through a body window.
One Laser System
This streamlined system uses a single multiphoton IR laser for imaging. SIM scanner for visible laser light stimulation is optional.
Dual Lines System
Employing the InSight DeepSee Dual laser, this system supplies dual wavelengths for multiphoton, multicolor imaging. SIM scanner for simultaneous laser light stimulation is also optional.
Twin Lasers System (with SIM Scanner)
This system employs two multiphoton IR lasers or two laser lines for imaging. In addition to multiphoton, multicolor imaging, simultaneous laser light stimulation is also supported in combination with an optional SIM scanner.
Lasers Adapted for a Range of Multiphoton Configurations
InSight DeepSee enables dual-wavelength simultaneous imaging for deep observation - with a high peak power with short 120fs pulse widths, a continuously variable broadband range from 680nm to 1300nm, and a fixed wavelength of 1040nm. A broad selection of dedicated models is available to make the most of multiphoton performance, including the MaiTai HP/eHP DeepSee-OL (Spectra-Physics)
|Spectra-Physics||MaiTai HP DeepSee-OL||690 nm - 1040 nm|
|MaiTai eHP DeepSee-OL||690 nm-1040 nm|
|InSight DS-OL||680 nm - 1300 nm|
|InSight DS Dual-OL||
680 nm - 1300 nm
1040 nm (fixed)
|COHERENT||Chameleon Vision I Olympus||690 nm - 1040 nm|
|Chameleon Vision II Olympus||680 nm - 1080 nm|
|Chameleon Vision S Olympus||690 nm - 1050 nm|
*Chameleon Series of Lasers not selling from Olympus in some region.
Laser Combiner for Visible Laser Light Stimulation
The laser combiner allows solid-state laser combinations for laser light stimulation at wavelengths of 405 nm, 458 nm, and 588 nm.
Light Guide Illumination Source U-HGLGPS
This light guide illumination source is equipped with a liquid light guide that minimizes the impact of vibration and lamp heat on the microscope and specimens alike. Employing a high-pressure mercury bulb, the light source offers a durable average lifetime of 2000 hours.
Transmitted Non-Descanned Light Detector
A high NA condenser and transmitted non-descanned light detector for multiphoton imaging detect fluorescence emitted from the focal plane and light scattered within the specimen.
Multialkali PMT 2CH Detector
Basic configuration of multialkali PMT 2CH.
Multialkali PMT 2CH + 2CH Detector
Multialkali PMT 2CH and optional addition of multialkali PMT 2CH.
Multialkali PMT 2CH + Cooled GaAsP PMT Detector
Multialkali PMT 2CH and optional cooled GaAsP PMT 2CH in combination.
|Laser Unit > IR Pulsed Laser with Negative Chirp for Multiphoton Excitation|
<Main IR pulsed laser>
|Laser Unit > Laser Combiner|
Introduction optic with AOM attenuation
|Scanning and Detection > Main Scanner > Standard Laser Ports|
|Scanning and Detection > Main Scanner > Detector > Standard|
|Scanning and Detection > Main Scanner > Detector > Cooled GaAsP-PMT 2 CH|
|Scanning and Detection > Main Scanner > Detector > Optional 4 CH|
|Scanning and Detection > Main Scanner > Photo Detection Method > Analog Integration|
|Scanning and Detection > Galvanometer Scanner (Normal Imaging) > Galvanometer Mirror Scanner (X, Y)|
|Scanning and Detection > Galvanometer Scanner (Normal Imaging) > Scanning Modes > 2D|
XY, XZ, XT, Xλ
|Scanning and Detection > Galvanometer Scanner (Normal Imaging) > Scanning Modes > 3D|
XYZ, XYT, XYλ, XZT, XTλ, XZλ
|Scanning and Detection > Galvanometer Scanner (Normal Imaging) > Scanning Modes > 4D|
XYZT, XZTλ, XYTλ
|Scanning and Detection > Galvanometer Scanner (Normal Imaging) > Scanning Modes > 5D|
|Scanning and Detection > Galvanometer Scanner (Normal Imaging) > Scanning Modes > Other|
ROI scanning: rectangle clip, ellipse, polygon, free area, line, free line and point
|Scanning and Detection > Galvanometer Scanner (Normal Imaging) > Scanning Speed|
|Scanning and Detection > Galvanometer Scanner (Normal Imaging) > Scanning Zoom|
|Scanning and Detection > Resonant Scanner (High-Speed Imaging) > Scanning Modes > 2D|
XY, XZ, XT
|Scanning and Detection > Resonant Scanner (High-Speed Imaging) > Scanning Modes > 3D|
XYZ, XYT, XZT
|Scanning and Detection > Resonant Scanner (High-Speed Imaging) > Scanning Modes > 4D|
|Scanning and Detection > Resonant Scanner (High-Speed Imaging) > Scanning Modes > 5D|
|Scanning and Detection > Resonant Scanner (High-Speed Imaging) > Scanning Modes > other|
ROI scanning: rectangle clip, line
|Scanning and Detection > Resonant Scanner (High-Speed Imaging) > Scanning Speed|
30 fps at 512 x 512, 438 fps at 512 x 32.
|Scanning and Detection > Resonant Scanner (High-Speed Imaging) > Scanning Zoom|
1.0 X - 8.0 X with 0.01 X increment
|Scanning and Detection > Field Number (NA)|
|Scanning and Detection > Z-Drive|
|Scanning and Detection > Transmitted Light Detector Unit|
Module with integrated external transmitted light photomultiplier detector and 100 W Halogen lamp, motorized switching, fi ber adaptation to microscope frame
|Microscope > Frame|
|System Control > Controller|
|System Control > Power Supply Unit|
|Optional Unit > SIM Scanner|
|Software > Basic Features|
Dark room matching GUI design. User-arrangeable layout.
|Software > IR Laser Controlling|
|Software > Optional Motorised-Stage Software|
XY motorised-stage control, map image acquisition for easy target locating. Tiling acquisition and software image stitching.
|Software > Optional Mapping and Multiplepoint Simulation Software|
Multiple-point stimulation and data acquisition software. Mapping multiple-point stimulation to generate reaction map. Filtering feature to select points.
|Software > Optional Sequencer Manager|
Advanced programmable software to define multiple imaging/ stimulation tasks and execute by hardware sequencer.
|Dimensions, Weight and Power Consumption > Microscope with Scan Unit > Dimensions (mm)|
|Dimensions, Weight and Power Consumption > Operating Environment (Indoor Use) > Ambient Temperature||Room temperature: 20 - 25 ℃|
|Dimensions, Weight and Power Consumption > Operating Environment (Indoor Use) > Maximum Relative Humidity||75 % or less at 25 °C requires continuous (24-hour) power supply|
4mm 3D Stack on Blood Vessel Label
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4mm 3D Stack on Blood Vessel Label with Texas Red in Mouse Brain
Image data courtesy of:
Hiroshi Hama, Rie Ito, Atsushi Miyawaki
Laboratory for Cell Function Dynamics, RIKEN Brain Science Institute