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
Two-photon optimized objectives- 16X (NA 0.8, WD 3 mm)
- 20X (NA 1.0, WD 2 mm)
- 25X (NA 1.10, WD 2 mm)
- 40X* (NA 0.8, WD 3.5 mm)
- 60X* (NA 1.0, WD 2.8 mm)
* Please note that these objectives have a smaller back aperture and cannot be used with the Axicon module
Nosepiece- Single position (M25 x 0.75 ou M32 x 0.75)
- Dual nosepiece designed to accomodate 1 two-photon optimized objective (M25 or M32) and 1 standard objective (RMS). Includes a DIC pocket. Easy adjustment of parfocality and XY alignment. Ideal for combining patch clamp and two-photon imaging

EXCITATION

Scanning Method16 facets polygonal scanner (x axis), matched with galvanometer mirror (y axis)
Polygonal scanning12 KHz polygonal scanner speed; 22 fps at 1024 x 512, dwell time per pixel of 83 ns/px, user-definable zoom up to 4x, frictionless rotation on air bearings
Laser power modulationPBS/half wave plate and high-speed grating modulator
Photostimulation/optogeneticsVisible and/or two-photon photostimulation; multi-point and user-adjustable region size, flash and tornado modes

SIGNAL DETECTION

Non-Descanned Detection (NDD, two-photon)Choice of 1-4 detectors: High-sensitivity Multi-Alkali PMTs, GaAsPs or SiPMs
Descanned Detectors for confocal add-onChoice of 1-4 detectors: High-sensitivity Multi-Alkali PMTs, GaAsPs or SiPMs
FLIM/PLIM Detectors1-2 photon counting sensitive fast photomultiplier tubes
Second and Third Harmonic Generation (SHG, THG)Choice of lasers and filters
CameraChoice of C-mount cameras
TrinocularTilting binocular tube with camera port and 10X eyepieces

LIGHT SOURCES

Two-photon laserMultiphoton laser input from 690 - 1300 nm. Integration of 1-4 lasers, Ti:sapphire or fiber lasers
Visible laserVisible fiber lasers and inputs for SPARQ (see "Extra Bliq modules" section), confocal, and photostimulation modules
EpifluorescenceMetal halide or LED epifluorescence light source
Transmitted lightHalogen or LED Illumination
Transmitted light modalitiesVisible and IR DIC, Transmitted NDD, Dodt gradient contrast, oblique illumination, TIRF, patterned photostimulation

MOTOR CONTROL

X,Y stageHeight-adjustable motorized XY bridge-stage, travel 65 mm x 65 mm, resolution: 20 nm, speed: 15 mm/sec. Possibility of integrating other manual or motorized stages
Z-FocusTravel range: 38 mm, resolution: 0.1 μm
Objective piezo-positioning systemZ travel: 500 μm, resolution: 0.5 nm
Depth of field control and bypass for AxiconMotorized depth of field adjustement from 5-40 μm with the Axicon (see "Extra Bliq modules" section). Includes a motorized bypass for standard Gaussian imaging

SOFTWARE AND COMPUTER

Nirvana ImagingSoftware fully integrated with scan head for easy imaging; customizable scan parameters for optimization of specimen excitation; integrated control of laser power and PMT high voltages; GPU computing for real-time analysis of data (intensity plots) while the images are acquired at full speed
X, Y, Z-moduleEasy creation of depth stacks and acquisition of 2D and 3D maps with user-customizable number of slices, step size and laser power
Multi-dimensional imagingEasy creation of complex series involving multiple dimensions: X, Y, Z, λ, t
Photostimulation/optogeneticsUser-definable points and region size, flash and tornado modes with synchronized laser modulation
Intensity plotsIntensity mapping and plotting of user-defined regions over time
Voltage outputsSignal outputs for electrophysiological experiments, stimulus control, and synchronization with external devices
ComputerApple Mac Mini with M2 Pro chip: 12 core CPU, 19 core GPU, 16 neural engine core, 32 Go of unified memory, 2 To of SSD storage, 8 To of additional storage and gigabit Ethernet, 27" 5K monitor

EXTRA BLIQ MODULES

Axicon: Optical Bessel beam volumetric imagingAxicon-based, self-healing non-diffractive Bessel beam imaging; user-definable depth of field from 5-40 μm; diffraction limited lateral resolution
SPARQ: Fast widefield optical sectioningSPARQ scrambling unit with up to 7 visible single-mode fiber lasers; speed of 10 fps; choice of camera. SPARQ control is supported byt the following software platforms: Nirvana (Mac), Inscoper, VisiView and Micro-Manager 2.0
Mouse holdersChoice of different stereotaxic chambers for small animals; head plate, ear and teeth bars, lung imaging, treadmill or custom
CustomizationAnisotropy, bone ablation, awake animals, grin endoscopy and other customizations available on request

 

* OPTIONS

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