LCS SPIM > LUXENDO

The Large Cleared Sample Light-Sheet Microscope

The Light-Sheet Microscope for Large Cleared Samples

Key Features

* High-performance, cost-effective bench-top device

* Unique cuvette-based sample mounting approach

* Fast 3D imaging of large cleared samples

* Compact, vibration-free and robust design

* Resolution down to 1 µm in 3D

* Browser-based user interface

 

Product Overview

The LCS SPIM is the latest product in Luxendo’s growing family of light-sheet microscopes. The basic configuration of the Luxendo LCS offers a cost-effective solution for cleared sample imaging. Based on a very versatile layout, the highly flexible upgrade options expand the performance of the system to increase speed and optical performance.

A quartz crystal cuvette, available in different lengths, facilitates mounting of large, delicate samples. The specimen is simply introduced into the cuvette filled with the clearing solution. The motorized sample stage for positioning and stack acquisition as well as the programmable optics concept for fast 3D scanning of the light-sheet through the sample ensure that the sample remains unperturbed during the imaging experiment while achieving the highest acquisition speed.

Typical applications of the microscope include imaging of large cleared-tissue samples, e.g. whole mouse brains to visualize complete neuronal and vascular networks. Microscopy of cleared organs or thick tissue samples is highly relevant in developmental biology studies and for cancer research, e.g. when investigating organ development, tumor structure and tumorigenesis.

Sample Chamber

An exchangeable quartz-crystal cuvette simplifies the whole sample mounting procedure as well as cleaning and maintenance, while ensuring compatibility with a wide range of clearing solutions by covering a refractive-index (RI) range from 1.33 to 1.57.

The cuvette is available in different lengths to accommodate samples of different sizes: the maximum cuvette dimensions are 40 mm × 20 mm × 20 mm (l × w × h).

Device Design

The LCS SPIM's compact, robust and vibration-free design provides maximal stability even during long-term high-throughput experiments.

Tailored to fit your lab bench, this class 1 laser system does not require any air table or vibration-compensation mechanism as all moving components are light-weight and balanced. Maximal stability of focus and thermal conditions are also guaranteed. The proprietary piezo-crawler stages ensure longevity and precision for a permanently accurate specimen positioning. Neither the images nor the quality of your sample are affected thanks to the unique and very gentle image-acquisition concept.

Illumination and Detection

The illumination optics comprise light-sheet generation by scanning a beam after passing a beam-shaper unit for length and diameter adjustment. Two Nikon 4x 0.2 NA air objective lenses project two aligned light sheets from opposing directions on the sample. The expandable and upgradable illumination setup allows a very flexible choice of lasers.

The detection optics comprises one Olympus 4x 0.28NA air/dry lens. An additional magnification changer results in a total magnification between 2x and 8x. The spatial detection arm is equipped with 10 position high-speed filter wheel. The robust sCMOS camera is well-suited for experiments that require high detection efficiency, quantification and speed.

Illumination	Detection	Effective Magnification	Field of View	Pixel Size	Optical Resolution
4x/0.2NA	Olympus 4x/0.28NA	2x	6600 µm	3.25 µm	1.1 µm
		4x	3300 µm	1.63 µm
		8x	1650 µm	0.81 µm

 

Laser combiner

* Maximum of 6 laser wavelengths (405, 445, 488, 515, 532, 561, 594, 642, and 685 nm with 40 mW each before fiber), more options available upon request

* Fast modulation (MHz range) and high extinction ratio

 

Illumination optics

* Chromatic correction from 440 to 660 nm

* Light-sheet generation by beam scanning

* Flexible light-sheet thickness

* 2 Nikon 4x 0.2 NA air objective lenses

* 2 identical illumination arms

* Optionally: axial beam scanning for an extended FOV

 

Detection optics

* One Olympus 4x 0.28 NA air/dry objective lens

* One detection arm equipped with a fast filter wheel (10 positions and 50 ms switching time between adjacent positions)

* Filters adapted to the selected laser lines

* One high-speed sCMOS camera

* Maximum frame rate up to 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% @ 560 nm

* Speed: 5 min for a whole mouse brain (12 mm x 12 mm x 12 mm)

 - 30s per stack

 - 4 stacks

 - Sampling size: 3 μm × 3 μm

Fast imaging mode

The basic configuration of the LCS SPIM can be upgraded with axial illumination scanning to extend the field of view (FOV) and to reduce shadowing effects.

Specifications

* Electronics, microscope software & computer

* Embedded microscope software with an open communication interface: documented API, TCP/IP-based communication

* Flexible GUI for interactive microscope control and experiment design

* Computer with 128GB RAM, Intel dual 8 core CPU

* High-speed RAID controller for data streaming, 8 × 4 TB local storage in RAID O

* GeForce RTX 2080 graphics card

* 10 Gbit/s on-board Ethernet port

* 1 UHTV 40 inch display

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