Multiphoton Video-rate Microscopy System (VMS)

Modular multiphoton microscope optimized for in vivo experiments

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Rolling myeloid cells (green, AlexaFluor647-anti-Ly6G) near the optic nerve head in the retina of a mouse after induction of inflammation. Contrast of vasculature and the retinal surface (blue) was obtained from reflectance.

Rolling myeloid cells (green, AlexaFluor647-anti-Ly6G) in the pial vessels of the meninges of a mouse in a model of meningitis. Vasculature (red) was labeled with FITC-dextran.

Both movies were acquired with a Video-Rate Microscopy System using a polygonal scanner. Courtesy of Clemens Alt Ph.D., Jean Carlos Cruz Hernandez Ph.D. and Charles P. Lin Ph.D, MGH Center for Systems Biology, Harvard University.

Both movies were acquired with a VMS using a polygonal scanner. Courtesy of Clemens Alt Ph.D., Jean Carlos Cruz Hernandez Ph.D. and Charles P. Lin Ph.D, MGH Center for Systems Biology, Harvard University.

High quality

Optimize the balance between frame rate and S/N ratio

VMS features a high speed polygonal scanner with an adjustable scan frequency from 9 KHz to 18 KHz enabling to set the frame rate from 15 to 30 fps. It uniquely allows you to find the optimal balance of S/N ratio and temporal resolution for each experiment.

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typical framerate and dwell time of polygonal and resonant scanner

The precise linearity of the polygonal scanner results in more time spent at each pixel (dwell time), more photons collected, and higher S/N ratio.

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polygonal scanner vs resonant scanner
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High speed

Explore a broad range of applications

With its speed of 32 to 2000 fps (1024×512 and 1024×6, respectively), the VMS is great for calcium imaging, dynamic cellular behavior, migration under flow and optogenetics. VMS technology also supports label-free techniques, including Coherent Anti-Stokes Raman Scattering (CARS), Second Harmonic Generation (SHG) and Third Harmonic Generation (THG).

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In vivo imaging of mouse GFP-labeled microglial cells (green) and myelin (blue, reflectance) before (left) and after (right) laser lesion (in red) . Movie: 30 minutes. Courtesy of Dr Yves De Koninck, CERVO Research Center, Université Laval.

High flexibility

Use the 2P properties to precisely control the photo-stimulation locations in 3D with low phototoxicity

Moreover, the uniform direction and rate of the polygonal scanner design result in highly precise synchronization of detection and stimulation events during imaging.

Calcium imaging widescreen

Image shows GCaMP6s fluorescence in mouse dorsal root ganglia (DRG) neurons after a paw stimulation. Scale bar: 50 um. Image courtesy of Dr Daniel C. Côté and Dr Yves De Koninck, CERVO Research Center, Université Laval.

bliq multi photon microscopy modules

High modularity

Choose between our multiple modules and options to personalize your VMS

Our systems are fully configurable. You can add our patented super-resolution, volumetric and optical sectioning modules. You can as well choose among many options such as photo-stimulation, confocal, FLIM, PLIM, GaAsP detectors, stereotaxic chambers, etc.

Specifications

GENERAL

FrameUpright, inverted and Open-Stand
2P objectives- 16X (NA 0.8, WD 3mm)
- 20X (NA 1.0, WD 2mm)
- 25X (NA 1.10, WD 2mm)
- 40X* (NA 0.8, WD 3.3mm)
- 60X* (NA 1.0, WD 2mm)
* Please note that these objectives have a smaller back aperture and cannot be used with our AXICON nor SLAM modules
Nosepiece- Single position (M25 x 0.75 ou M32 x 0.75)
- Dual nosepiece designed to accommodate one 2P optimized objective (M25 ou M32) and one standard objective (RMS). Ideal for combining 2P imaging and electrophysiology

EXCITATION

Scanning Method36 facets Polygonal scanner matched with galvanometer mirror
Scan Speed32fps at 1024 x512 and up to 2000fps at 1024 x 6, pixel size max: 1024 x 1024
Polygonal scanner speedPolygonal scanner speed modulation from 9KHz to 18KHz; adjustable dwell time per pixel from 50ns/px to 100ns/px
Scan Customization User-definable optical zoom up to 8x; user-definable pixels per line and lines per scan from 6-1024; user-definable frame rate up to 2000fps
Laser power controlPBS and half wave plate, AOM and EOM
Line scan imaging *Second set of matched galvanometers to provide line scan capabilites; point scan; 360° of line scan rotation
Photostimulation *Visible and/or multiphoton photostimulation; multi-points and user adjustable region size. Choice of visible and IR lasers

SIGNAL DETECTION

Non-Descanned Detectors (2P)Choice of 1-3 detectors: High-sensitivity Hamamatsu Multi-Alkali PMTs or GaAsPs. Upgradeable to up to 6 detectors
Descanned reflectance *Contrast detection system allowing for descan contrast imaging using an APD
Descanned Detectors for confocal add-on *Choice of 1-3 detectors: Multi-Alkali PMTs or GaAsPs
FLIM Detectors *1-2 single photon sensitive fast photomultiplier tubes; thermoelectric cooling and automatic overload protection available as an option
Second and Third Harmonic Generation (SHG, THG)Choice of filters
Camera *Standard C-mount camera port built into scan head. Choice of camera

OPTICAL INPUTS

Multiphoton laserOptimized for multiphoton laser input from 690 - 1700 nm. Integration of 1-4 multiphoton lasers
Visible laser *Fiber laser inputs for widefield optical sectioning (SPARQ), confocal add-on or photostimulation
Epifluorescence *Metalhalyde or LED epifluorescence light source
Transmitted light *Visible and IR DIC; halogen or LED Illumination

MOTOR CONTROL

X,Y stageHeight-adjustable motorized XY bridge-stage, travel 65mm x 65mm, resolution 20 nanometers, speed 15mm/sec
Z-FocusTravel range of 38 mm with ~0.1 μm step size
Z-Piezo *Travel range of up to 500 μm with 0.05 μm step size
Depth of field control for Axicon *Motorized depth of field adjustement up to 50um with 5um step size, with the AXICON
Simultaneous region size stimulation *User-selectable ROI stimulation from 30um to 120um with 30um step size

SOFTWARE AND COMPUTER

Nirvana ImagingSoftware fully integrated with scan head for easy imaging; customizable scan settings for optimization of specimen excitation; integrated control of laser power and PMT high voltages
X,Y,Z-moduleEasy creation of depth stacks and acquisition of 2D and 3D maps with user-customizable slice number, step size and laser power
T-moduleEasy creation of complex series involving X,Y,Z-modules and triggered options
PhotostimulationUser-defined points and regions with synchronized laser modulation
Regions of InterestIntensity mapping and plotting for user-defined regions over time
Voltage Inputs/OutputsSignal inputs and outputs for electrophysiological experiments, stimulus control, and synchronization with external devices
Apple computer4.2GHz, 64GB SDRAM, 2TB SSD, 48TB extra storage, 27" 5K monitor

BLIQ UNIQUE MODULES *

Optical Bessel beam volumetric imaging: AXICONUser-definable depth of field from 10um to 50um; diffraction limited lateral resolution. Does not affect the scan speed
Fast widefield optical sectioning: SPARQUp to 7 visible fibered lasers; Speed up to 10fps; sCMOS 16-bit camera, 2048 x 2048 pixel array
Super-resolution for 2P and confocal (SLAM)High speed super-resolution module down to 120 nm XY resolution; Scan speed 1024 x 512 @ 16 fps
Dual nosepiece for 2P and standard imagingFits 1 large back aperture objective and 1 standard RMS objective. Each position has a DIC pocket. Low movement vibration. Easy adjustement of parfocality and XY alignment. No tools required
Mouse holdersChoice of different stereotaxic chambers for small animals; head plate, ear and teeth bars, lung imaging, treadmill or custom
Trigger box16 BNC ports: 4 analog inputs, 4 analog outputs, 4 digital inputs and 4 digital outputs

 

* OPTIONS

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