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Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours

Imaging living organisms at high spatial resolution requires effective and innocuous immobilization. Long-term imaging places further demands on sample mounting with minimal perturbation of the organism. Here we present a simple, inexpensive method for rapid encapsulation of small animals of any dev...

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Autores principales: Burnett, Kyra, Edsinger, Eric, Albrecht, Dirk R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6123791/
https://www.ncbi.nlm.nih.gov/pubmed/30271954
http://dx.doi.org/10.1038/s42003-018-0079-6
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author Burnett, Kyra
Edsinger, Eric
Albrecht, Dirk R.
author_facet Burnett, Kyra
Edsinger, Eric
Albrecht, Dirk R.
author_sort Burnett, Kyra
collection PubMed
description Imaging living organisms at high spatial resolution requires effective and innocuous immobilization. Long-term imaging places further demands on sample mounting with minimal perturbation of the organism. Here we present a simple, inexpensive method for rapid encapsulation of small animals of any developmental stage within a photo-crosslinked polyethylene glycol (PEG) hydrogel, gently restricting movement within their confined spaces. Immobilized animals maintain their original morphology in a hydrated environment compatible with chemical treatment, optical stimulation, and light-sheet microscopy. We demonstrate prolonged three-dimensional imaging of neural responses in the nematode Caenorhabditis elegans, recovery of viable organisms after 24 h, and imaging of larger squid hatchlings. We characterize a range of hydrogel and illumination conditions for immobilization quality, and identify paralytic-free conditions suitable for high-resolution single-cell imaging. Overall, PEG hydrogel encapsulation provides fast, versatile, and gentle mounting of small living organisms, from yeast to zebrafish, for continuous observation over hours.
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spelling pubmed-61237912018-09-28 Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours Burnett, Kyra Edsinger, Eric Albrecht, Dirk R. Commun Biol Article Imaging living organisms at high spatial resolution requires effective and innocuous immobilization. Long-term imaging places further demands on sample mounting with minimal perturbation of the organism. Here we present a simple, inexpensive method for rapid encapsulation of small animals of any developmental stage within a photo-crosslinked polyethylene glycol (PEG) hydrogel, gently restricting movement within their confined spaces. Immobilized animals maintain their original morphology in a hydrated environment compatible with chemical treatment, optical stimulation, and light-sheet microscopy. We demonstrate prolonged three-dimensional imaging of neural responses in the nematode Caenorhabditis elegans, recovery of viable organisms after 24 h, and imaging of larger squid hatchlings. We characterize a range of hydrogel and illumination conditions for immobilization quality, and identify paralytic-free conditions suitable for high-resolution single-cell imaging. Overall, PEG hydrogel encapsulation provides fast, versatile, and gentle mounting of small living organisms, from yeast to zebrafish, for continuous observation over hours. Nature Publishing Group UK 2018-06-21 /pmc/articles/PMC6123791/ /pubmed/30271954 http://dx.doi.org/10.1038/s42003-018-0079-6 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Burnett, Kyra
Edsinger, Eric
Albrecht, Dirk R.
Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours
title Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours
title_full Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours
title_fullStr Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours
title_full_unstemmed Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours
title_short Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours
title_sort rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6123791/
https://www.ncbi.nlm.nih.gov/pubmed/30271954
http://dx.doi.org/10.1038/s42003-018-0079-6
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