Multiphoton Bessel beam
light sheet system

Image courtesy of Dr. Martin Lévesque, CERVO Research Center, Université Laval

Unique

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.

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bessel light sheet

Bliq's Bessel light sheet system. © Cléophace Akitegetse, Université Laval, 2018

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bessel

Diagram of the Bessel focal line. © Cléophace Akitegetse, Université Laval, 2018

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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.

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Dopaminergic neurons stained with Tyrosine Hydroxylase in iDisco cleared P1 mouse brain. Courtesy of Dr. Martin Lévesque, CERVO Research Center, Université Laval

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obstruccion bessel

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

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Low photodamage

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.

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obstruccion bessel

MIP of desmine-594 expression in a iDISCO cleared mouse heart. © Cléophace Akitegetse, Université Laval, 2018

High quality

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.

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obstruccion bessel

Blood vessels stained with CD31 in cleared adult mouse cortex. Courtesy of Dr. Martin Lévesque and Dr. Caroline Ménard, CERVO Research Center, Université Laval

Personalized

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.

Specifications

Detection objectives

10X: NA 0.6, WD 8mm
20X: NA 0.95, WD 9.9mm

Resolution (10X)


- Lateral: 650nm
- Axial: 2μm

FOV (10X)

1.3mm x 1.3mm

Acquisition speed per FOV

Up to 100FPS (10ms exposure time)

Chamber size

6cm x 4cm x 3cm. Custom chambers also available

Sample size

Up to 40mm x 100mm x 7mm (without rotation of specimen)

Detection

- 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

Incubation media

Hydrogel, aqueous and solvent-based methods (CLARITY, Scale, CUBIC, iDisco, DBE, etc.)