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

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 imag...

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Autores principales: Smith, Jayson J., Kenny, Isabel W., Wolff, Carsten, Cray, Rachel, Kumar, Abhishek, Sherwood, David R., Matus, David Q.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Cell and Developmental Biology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9586288/
https://www.ncbi.nlm.nih.gov/pubmed/36274853
http://dx.doi.org/10.3389/fcell.2022.1012820
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author Smith, Jayson J.
Kenny, Isabel W.
Wolff, Carsten
Cray, Rachel
Kumar, Abhishek
Sherwood, David R.
Matus, David Q.
author_facet Smith, Jayson J.
Kenny, Isabel W.
Wolff, Carsten
Cray, Rachel
Kumar, Abhishek
Sherwood, David R.
Matus, David Q.
author_sort Smith, Jayson J.
collection PubMed
description 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.
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spelling pubmed-95862882022-10-22 A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults Smith, Jayson J. Kenny, Isabel W. Wolff, Carsten Cray, Rachel Kumar, Abhishek Sherwood, David R. Matus, David Q. Front Cell Dev Biol Cell and Developmental Biology 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. Frontiers Media S.A. 2022-10-07 /pmc/articles/PMC9586288/ /pubmed/36274853 http://dx.doi.org/10.3389/fcell.2022.1012820 Text en Copyright © 2022 Smith, Kenny, Wolff, Cray, Kumar, Sherwood and Matus. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Smith, Jayson J.
Kenny, Isabel W.
Wolff, Carsten
Cray, Rachel
Kumar, Abhishek
Sherwood, David R.
Matus, David Q.
A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults
title A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults
title_full A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults
title_fullStr A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults
title_full_unstemmed A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults
title_short A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults
title_sort light sheet fluorescence microscopy protocol for caenorhabditis elegans larvae and adults
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9586288/
https://www.ncbi.nlm.nih.gov/pubmed/36274853
http://dx.doi.org/10.3389/fcell.2022.1012820
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