Nature methods. 2022 May 11. doi: 10.1038/s41592-022-01468-5. pii: 10.1038/s41592-022-01468-5 |
A hybrid open-top light-sheet microscope for versatile multi-scale imaging of cleared tissues. |
Glaser AK1, Bishop KW2, Barner LA3, Susaki EA4, Kubota SI5, Gao G6, Serafin RB7, Balaram P8, Turschak E9, Nicovich PR10, Lai H11, Lucas LAG12, Yi Y13, Nichols EK14, Huang H15, Reder NP16, Wilson JJ17, Sivakumar R18, Shamskhou E19, Stoltzfus CR20, Wei X21, Hempton AK22, Pende M23, Murawala P24, Dodt HU25, Imaizumi T26, Shendure J27, Beliveau BJ28, Gerner MY29, Xin L30, Zhao H31, True LD32, Reid RC33, Chandrashekar J34, Ueda HR35, Svoboda K36, Liu JTC37 |
Abstract 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. |
© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc. |
Publikations ID: 35545715 Quelle: öffnen |