MuVi-SPIM_CS > LUXENDO

The Multiple View Light-Sheet Microscope for Cleared Samples

3D imaging of optically cleared samples

The LUXENDO MuVi SPIM CS is an extension of the MuVi SPIM capabilities to enable 3D imaging of optically cleared samples.

The sample chamber, the illumination and the detection unit in the octagon are adapted to match the experimental needs in terms of sample size, refractive index, field-of-view and resolution, adding high flexibility to the system.

 

 

Cleared Sample Module

The Cleared Sample Module is an add-on consisting of an exchangeable cleared sample octagon and an overhead sample positioning unit installed on the microscope’s top plate.

The octagon can be easily removed from the microscope for maintenance and cleaning of the sample chamber and the objective lenses that have been exposed to the mounting solution. The sample chamber is compatible with a broad range of solutions used in different clearing protocols. Available in two different sizes (max. volumes of 20 ml and 33 ml), the sample chamber can accommodate large samples such as mouse brains and other complete organs. Sample mounting from above enables easy sample access and optimized travel range suited for various mounting methods (e.g. hook, plate, pin or cuvette).

Flexible illumination and detection configurations leverage the MuVi SPIM CS to meet your application needs and allow imaging of a great variety of samples at high speed and high resolution. 

 

Illumination and Detection 

The MuVi SPIM CS (Cleared Sample) can achieve a resolution down to 300 nm in 3D (at a wavelength of 500 nm), enabling imaging of optically cleared samples.

Two Nikon 10x 0.3 NA air objective lenses project two aligned light sheets from opposing directions on the sample. Detection includes high numerical aperture, multi-immersion objective lenses, with 20x and 10x magnification. An additional magnification changer results in a total magnification that ranges from 7.5x to 30x. You can choose the set of illumination and detection objective lenses according to the experimental needs.

Illumination		Detection		Effective Magnification	Field of View	Pixel size	Optical Resolution
10x/0.3 NA		Olympus 20x/1.0 NA		15.0x	890 µm	434 nm	300 nm
				20.0x	665 µm	325 nm
				30.0x	445 µm	217 nm
10x/0.3 NA		Nikon 10x/0.5 NA		7.5x	1775 µm	867 nm	600 nm
				10.0x	1330 µm	650 nm
				15.0x	890 µm	434 nm

 

Illumination Optics

• Chromatic correction from 440 to 660 nm
• Light-sheet generation by beam scanning
• Flexible light-sheet thickness (2 µm to 8 µm)
• 2 Nikon 10x 0.3 NA air objective lenses
• 2 identical illumination arms

 

Detection Optics

• Different combinations of lenses possible:

• 1 Nikon 20x 1.0 NA 8 mm WD multi-immersion

• 1 Nikon 10x 0.5 NA 5.5 mm WD

• Combinations of 2 of them
• 1 or 2 detection arms, each equipped with a fast filter wheel (10 positions and 50 ms switching time between adjacent positions)
• Filters adapted to the selected laser lines
• 2 high-speed sCMOS cameras Hamamatsu Orca Flash 4.0 V3
• Maximum frame rate >80 fps at full frame (2048 × 2048 pixels of 6.5 µm × 6.5 µm size) and up to 500 fps at subframe cropping
• Peak quantum efficiency (QE): 82% @ 560nm

 

Sample Chamber

Single-sided 10x detection:

30 mm (lateral) × 14 mm (depth) × 40 mm (height)

Single-sided 20x detection:

30 mm (lateral) × 14 mm (depth) × 40 mm (height)

Dual-sided 10x detection:

30 mm (lateral) × 11.6 mm (depth) × 40 mm (height)

Dual-sided 20x detection:

30 mm (lateral) × 16.4 mm (depth) × 40 mm (height)

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