A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults

Jayson J. Smith; Jayson J. Smith; Jayson J. Smith; Isabel W. Kenny; Isabel W. Kenny; Carsten Wolff; Carsten Wolff; Rachel Cray; Abhishek Kumar; Abhishek Kumar; David R. Sherwood; David R. Sherwood; David Q. Matus; David Q. Matus

doi:10.3389/fcell.2022.1012820

Frontiers in Cell and Developmental Biology (Oct 2022)

A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults

Jayson J. Smith,
Jayson J. Smith,
Jayson J. Smith,
Isabel W. Kenny,
Isabel W. Kenny,
Carsten Wolff,
Carsten Wolff,
Rachel Cray,
Abhishek Kumar,
Abhishek Kumar,
David R. Sherwood,
David R. Sherwood,
David Q. Matus,
David Q. Matus

Affiliations

Jayson J. Smith: Department of Neurobiology, University of Chicago, Chicago, IL, United States
Jayson J. Smith: University of Chicago Neuroscience Institute, Chicago, IL, United States
Jayson J. Smith: Embryology: Modern Concepts and Techniques, Marine Biological Laboratory, Woods Hole, MA, United States
Isabel W. Kenny: Embryology: Modern Concepts and Techniques, Marine Biological Laboratory, Woods Hole, MA, United States
Isabel W. Kenny: Department of Biology, Duke University, Durham, NC, United States
Carsten Wolff: Embryology: Modern Concepts and Techniques, Marine Biological Laboratory, Woods Hole, MA, United States
Carsten Wolff: Marine Biological Laboratory, Woods Hole, MA, United States
Rachel Cray: Marine Biological Laboratory, Woods Hole, MA, United States
Abhishek Kumar: Embryology: Modern Concepts and Techniques, Marine Biological Laboratory, Woods Hole, MA, United States
Abhishek Kumar: Marine Biological Laboratory, Woods Hole, MA, United States
David R. Sherwood: Embryology: Modern Concepts and Techniques, Marine Biological Laboratory, Woods Hole, MA, United States
David R. Sherwood: Department of Biology, Duke University, Durham, NC, United States
David Q. Matus: Embryology: Modern Concepts and Techniques, Marine Biological Laboratory, Woods Hole, MA, United States
David Q. Matus: Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, NY, United States

DOI: https://doi.org/10.3389/fcell.2022.1012820
Journal volume & issue: Vol. 10

Abstract

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Light sheet fluorescence microscopy (LSFM) has become a method of choice for live imaging because of its fast acquisition and reduced photobleaching and phototoxicity. Despite the strengths and growing availability of LSFM systems, no generalized LSFM mounting protocol has been adapted for live imaging of post-embryonic stages of C. elegans. A major challenge has been to develop methods to limit animal movement using a mounting media that matches the refractive index of the optical system. Here, we describe a simple mounting and immobilization protocol using a refractive-index matched UV-curable hydrogel within fluorinated ethylene propylene (FEP) tubes for efficient and reliable imaging of larval and adult C. elegans stages.

Published in Frontiers in Cell and Developmental Biology

ISSN: 2296-634X (Online)
Publisher: Frontiers Media S.A.
Country of publisher: Switzerland
LCC subjects: Science: Biology (General)
Website: https://www.frontiersin.org/journals/cell-and-developmental-biology

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