Multi-line Lasers Help Simplify Light Sheet Microscopes Targeting Clinical Applications

Session

Poster Session

Authors

Melissa Haahr (1), Magnus Rådmark (1), Steven Edwards (3), Kohei Otomo (2), Etsuo A. Susaki (2)

Affiliations

1. Cobolt AB, a part of Hübner Photonics
2. Department of Biochemistry and Systems Biomedicine, Juntendo University Graduate School of Medicine
3. SciLifeLab, Department of Applied Physics, Royal Institute of Technology

Keywords

Multi-line laser, light sheet microscopy, fluorescence microscopy, lasers for microscopy, descSPIM, clinical diagnostics, bio-imaging

Abstract text

Fluorescence imaging using lasers is a key technique in both biomedical research as well as in clinical applications. Fluorescence based microscopy for high-resolution and high-content imaging typically relies on the use of several individual laser sources at different wavelengths. This is often solved by using a laser-combiner, which includes separate lasers and beam-combining optics. Although flexible, laser combiners are usually bulky solutions that are difficult to keep aligned. A simpler solution for stable, simple and service-free integration of multi-color illumination is to use a permanently aligned multi-line laser.

Multi-line lasers are perfectly suited for bringing fluorescence imaging techniques into clinical settings, where reliability, repeatability and user-friendliness are prerequisites, but which still require high resolution images with high level of molecular specificity for precise analysis. Light sheet microscopy is one such fluorescence imaging method that has demonstrated particularly strong capability to provide new ways of analyzing human specimens in the field of pathology and offering digital alternatives to the decades old and well-established field of histopathology [1]. Light sheet microscopy brings the combination of gentle sample illumination, high speed 3D imaging as well as high spatial resolution and molecular specificity.

In this work we demonstrate how the concept of multi-color light sheet microscopy can be further improved in terms of simplicity and flexibility through the combination of descSPIM (desktop equipped Selective Plane Illumination Microscopy) with new multi-line laser sources. The descSPIM offers the benefits of both efficient sample evaluation in routine experimentation, with volume imaging of optically cleared samples in minutes, as well as advanced capabilities, including multi-view whole-brain imaging and the assessment of antibody drug distribution in a tumor xenograft model [2]. Using a multi-line laser as the illumination source enables ultimately compact, simple and flexible system integration. This permanently aligned multi-line laser can be customized with any combination of more than 14 colors, ranging from 405 nm to 660 nm. As a further enhancement of the system flexibility, we will also demonstrate the introduction of a new compact and directly modulated laser line in the orange (594 nm), specifically suitable for excitation of mCherry and other red fluorescent proteins. 

References

[1] Glaser, A. K. et al. Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens. Nat. Biomed. Eng. 1, 0084 (2017).

[2] Otomo K et.al Biorxiv descSPIM: Affordable and Easy-to-Build Light-Sheet Microscopy for Tissue Clearing Technique Users.