A hybrid open-top light-sheet microscope for versatile multi-scale imaging of cleared tissues

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 transp...

Full description

Saved in:
Bibliographic Details
Published inNature methods Vol. 19; no. 5; pp. 613 - 619
Main Authors Glaser, Adam K., Bishop, Kevin W., Barner, Lindsey A., Susaki, Etsuo A., Kubota, Shimpei I., Gao, Gan, Serafin, Robert B., Balaram, Pooja, Turschak, Emily, Nicovich, Philip R., Lai, Hoyin, Lucas, Luciano A. G., Yi, Yating, Nichols, Eva K., Huang, Hongyi, Reder, Nicholas P., Wilson, Jasmine J., Sivakumar, Ramya, Shamskhou, Elya, Stoltzfus, Caleb R., Wei, Xing, Hempton, Andrew K., Pende, Marko, Murawala, Prayag, Dodt, Hans-Ulrich, Imaizumi, Takato, Shendure, Jay, Beliveau, Brian J., Gerner, Michael Y., Xin, Li, Zhao, Hu, True, Lawrence D., Reid, R. Clay, Chandrashekar, Jayaram, Ueda, Hiroki R., Svoboda, Karel, Liu, Jonathan T. C.
Format Journal Article
LanguageEnglish
Published New York Nature Publishing Group US 01.05.2022
Nature Publishing Group
Subjects
631/1647/245/2225
631/1647/328/2237
Animals
Axon guidance
Axons
Bioinformatics
Biological Microscopy
Biological Techniques
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Brain
Brain cancer
Fluorescence microscopy
Hybrid systems
Imaging
Life Sciences
Light sheets
Metastases
Metastasis
Mice
Microscopy
Microscopy - methods
Multiscale analysis
Neuroimaging
Proteomics
Sample holders
Tissues
Online AccessGet full text
ISSN1548-7091
1548-7105
1548-7105
DOI10.1038/s41592-022-01468-5

Cover

More Information
Summary: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. A ‘hybrid’ open-top light-sheet microscope is described, which can be used for broad multi-scale volumetric imaging of one or more large tissues, cleared with diverse protocols, and conveniently mounted on an array of sample holders.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 14
content type line 23
A.K.G. and J.T.C.L. conceived of and designed the microscope system. P.R.N. provided feedback on the system design and its potential applications. A.K.G., K.W.B., R.B.S., and G.G. performed simulations of the microscope. A.K.G. fabricated the microscope system with help from L.A.B. E.A.S. and H.R.U. provided and prepared the immunostained CUBIC-cleared mouse brains. E.A.S., S.I.K., and H.R.U. prepared and provided the metastatic mouse brains. J.C. and K.S. prepared and provided the mouse brain. P.B., E.T., and R.C.R. provided the mouse brain with preparation by A.K.G. The human brain slice was prepared by A.K.G. H.L. and L.A.G.L. provided quantitative analysis of the high-resolution images. Y.Y. and H.Z. provided and prepared the PEGASOS-cleared mouse brain. E.K.N., B.J.B., and J.S. provided and prepared the SHIELD-cleared mouse embryos. H.H., N.P.R., and L.D.T. provided and prepared the ECi-cleared human prostate tissue. J.J.W., R.S., E.S., C.R.S., and M.Y.G. provided and prepared the Ce3D-cleared mouse lymph node. X.W. and L.X. provided and prepared the iDISCO-cleared mouse prostate. A.K.H. and T.I. provided and prepared the ClearSee-processed Arabidopsis plant and M.P., P.M., and H.U.D. provided and prepared the DEEP-Clear-processed Axolotl. A.K.G. and J.T.C.L. led the writing of the manuscript. All authors contributed to the manuscript.
Author Contributions
ISSN:1548-7091
1548-7105
1548-7105
DOI:10.1038/s41592-022-01468-5