Open AccessCompassLSM: axially swept light-sheet microscopy made simple
Author(s) -
Yehe Liu,
Andrew M. Rollins,
Michael W. Jenkins
Publication year - 2021
Publication title -
biomedical optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.440292
Subject(s) - light sheet fluorescence microscopy , optical sectioning , optics , axial symmetry , chromatic aberration , computer science , microscopy , field of view , clearance , chromatic scale , physics , scanning confocal electron microscopy , medicine , quantum mechanics , urology
Axially swept light-sheet microscopy (ASLM) is an effective method of generating a uniform light sheet across a large field of view (FOV). However, current ASLM designs are more complicated than conventional light-sheet systems, limiting their adaptation in less experienced labs. By eliminating difficult-to-align components and reducing the total number of components, we show that high-performance ASLM can be accomplished much simpler than existing designs, requiring less expertise and effort to construct, align, and operate. Despite the high simplicity, our design achieved 3.5-µm uniform optical sectioning across a >6-mm FOV, surpassing existing light-sheet designs with similar optical sectioning. With well-corrected chromatic aberration, multi-channel fluorescence imaging can be performed without realignment. This manuscript provides a comprehensive tutorial on building the system and demonstrates the imaging performance with optically cleared whole-mount tissue samples.