Multiphoton Bessel beam
light sheet system
How does it work?
Light sheet microscopy is generally performed using a visible Gaussian beam scanned in parallel to a camera detector plane. Since the depth of field and the beam waist are closely related, the technology becomes challenging when there is a need to image bigger sample and increase the field of illumination (determined by the beam length), without compromising on the axial resolution (determined by the light sheet thickness).
To overcome this challenge, we use an Axicon lens to form thin needle-shaped beam (Bessel beam) which, when swept at a very high speed along an axis, forms a light sheet. The main advantage of this strategy resides in its capacity to create a light sheet of uniform thickness along its longitudinal profile and thus provide large volume images with an isotropic subcellular resolution.
Bliq's Bessel light sheet system. © Cléophace Akitegetse, Université Laval, 2018
Diagram of the Bessel focal line. © Cléophace Akitegetse, Université Laval, 2018
Image big samples
Obtain large FOV with subcellular resolution
Bliq’s Bessel beam light sheet system provides a FOV wide enough to image a section of a rodent’s brain while maintaining micrometer-level isotropic resolution. Furthermore, the illumination profile of a Bessel beam remains undisturbed when there is an obstruction within the sample which reduces shading compared to standard Gausian beam technology and ensures that no data is lost.
Dopaminergic neurons stained with Tyrosine Hydroxylase in iDisco cleared P1 mouse brain. Courtesy of Dr. Martin Lévesque, CERVO Research Center, Université Laval
Diagram of the resulting Bessel beam when using an axicon (A) and Bessel beam re-forming after obstruction (B). Ref.: Egmason // CC-BY-SA-3.0
Perform deep multiphoton light sheet imaging with low phototoxicity
Take advantage of all the benefits of multiphoton microscopy, including less photobleaching and photodamage as well as the removal of out-of-focus plane fluorescence.
MIP of desmine-594 expression in a iDISCO cleared mouse heart. © Cléophace Akitegetse, Université Laval, 2018
Achieve high axial resolution Bessel illumination
Bliq carefully selects high-quality Axicon lenses to generate a long, thin and uniform Bessel beam that provides up to 2μm axial resolution across the FOV.
Blood vessels stained with CD31 in cleared adult mouse cortex. Courtesy of Dr. Martin Lévesque and Dr. Caroline Ménard, CERVO Research Center, Université Laval
Use a variety of sample sizes with our large and custom chambers
With Bliq’s imaging chamber, you can image samples up to 40mm x 100mm x 7mm without having to rotate them. Our dedicated team is also available to design and produce custom chambers for your specific needs.
|10X: NA 0.6, WD 8mm
20X: NA 0.95, WD 9.9mm
|- Lateral: 650nm
- Axial: 2μm
|1.3mm x 1.3mm|
Acquisition speed per FOV
|Up to 100FPS (10ms exposure time)|
|6cm x 4cm x 3cm. Custom chambers also available|
|Up to 40mm x 100mm x 7mm (without rotation of specimen)|
|- Hamamatsu Orca Flash 4.0 100FPS, 2048 x 2048 (other options are also available)
- 1-channel or simultaneous 2-channel detection
Laser source: Ultrafast Amplified IR laser
|- 650nm to 900nm and 1200nm to 2500nm
- Output power: > 3W
- Pulse energy: >3μJ
- Pulse duration: <100fs
- Repetition rate: 1MHz
Software and computer
|- Bliq's Nirvana acquisition software
- Apple iMac Pro, 4,2GHz, 128GB SDRAM, 4TB SSD, VEGA 16GB video, 80TB of storage, 27″ 5K monitor
|Hydrogel, aqueous and solvent-based methods (CLARITY, Scale, CUBIC, iDisco, DBE, etc.)|