Published March 29, 2024 | Version v1
Journal article Open

Projective light-sheet microscopy with flexible parameter selection

Description

Projection imaging accelerates volumetric interrogation in fluorescence microscopy, but for multi-cellular samples, the resulting images may lack contrast, as many structures and haze are summed up. Here, we demonstrate rapid projective light-sheet imaging with parameter selection (props) of imaging depth, position and viewing angle. This allows us to selectively image different sub-volumes of a sample, rapidly switch between them and exclude background fluorescence. Here we demonstrate the power of props by functional imaging within distinct regions of the zebrafish brain, monitoring calcium firing inside muscle cells of moving Drosophila larvae, super-resolution imaging of selected cell layers, and by optically unwrapping the curved surface of a Drosophila embryo. We anticipate that props will accelerate volumetric interrogation, ranging from subcellular to mesoscopic scales.

Data availability

The image and movie data generated in this study have been deposited in a public Zenodo repository: https://zenodo.org/records/10182659 Source data are provided with this paper.

The Matlab and Python codes used for data analysis are available in a public GitHub repository. The LabView-based control software used within this manuscript to control the OPM and LLSM systems is freely available for nonprofit institutions through a Material Transfer Agreement with UT Southwestern Medical Center.

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Additional details

Identifiers

DOI
10.1038/s41467-024-46693-y
Other
oai:uchicago.tind.io:11466

Funding

National Institutes of Health
R35GM133522
National Institutes of Health
R35GM127091
National Institutes of Health
R01AR075005
National Institutes of Health
R01AR053173
National Institutes of Health
R35GM136316
National Institutes of Health
1R01GM134207
National Institutes of Health
1R21HD105189
Robert A Welch Foundation
I-1950-20180324

UChicago Information

Division(s)
Biological Sciences Division
Department(s)
Molecular Genetics and Cell Biology