Open AccessA hybrid open-top light-sheet microscope for versatile multi-scale imaging of cleared tissues
Author(s) -
Adam K. Glaser,
Kevin W. Bishop,
Lindsey A. Barner,
Etsuo A. Susaki,
Shimpei I. Kubota,
Gan Gao,
Robert Serafin,
Pooja Balaram,
Emily Turschak,
Philip R. Nicovich,
Hoyin Lai,
Luciano Lucas,
Yating Yi,
Eva Nichols,
Hongyi Huang,
Nicholas P. Reder,
Jasmine Wilson,
Ramya Sivakumar,
Elya Shamskhou,
Caleb Stoltzfus,
Xing Wei,
Andrew K Hempton,
Marko Pende,
Prayag Murawala,
HansUlrich Dodt,
Takato Imaizumi,
Jay Shendure,
Brian J. Beliveau,
Michael Y. Gerner,
Xin Li,
Hu Zhao,
Lawrence D. True,
R. Clay Reid,
Jayaram Chandrashekar,
Hiroki Ueda,
Karel Svoboda,
Jonathan Liu
Publication year - 2022
Publication title -
nature methods
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 19.469
H-Index - 318
eISSN - 1548-7105
pISSN - 1548-7091
DOI - 10.1038/s41592-022-01468-5
Subject(s) - clearance , light sheet fluorescence microscopy , microscopy , throughput , microscope , biomedical engineering , computer science , materials science , nanotechnology , scale (ratio) , optics , scanning confocal electron microscopy , medicine , telecommunications , physics , quantum mechanics , wireless , urology
Light-sheet microscopy has emerged as the preferred means for high-throughput volumetric imaging of cleared tissues. However, there is a need for a flexible system that can address imaging applications with varied requirements in terms of resolution, sample size, tissue-clearing protocol, and transparent sample-holder material. Here, we present a 'hybrid' system that combines a unique non-orthogonal dual-objective and conventional (orthogonal) open-top light-sheet (OTLS) architecture for versatile multi-scale volumetric imaging. We demonstrate efficient screening and targeted sub-micrometer imaging of sparse axons within an intact, cleared mouse brain. The same system enables high-throughput automated imaging of multiple specimens, as spotlighted by a quantitative multi-scale analysis of brain metastases. Compared with existing academic and commercial light-sheet microscopy systems, our hybrid OTLS system provides a unique combination of versatility and performance necessary to satisfy the diverse requirements of a growing number of cleared-tissue imaging applications.