Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying.

The cellular cortex is an approximately 200-nm-thick actin network that lies just beneath the cell membrane. It is responsible for the mechanical properties of cells, and as such, it is involved in many cellular processes, including cell migration and cellular interactions with the environment. To d...

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Autores principales:	Moritz Schu, Emmanuel Terriac, Marcus Koch, Stephan Paschke, Franziska Lautenschläger, Daniel A D Flormann
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fb456fcc79dd48cfb019c88a7a8a1c22
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spelling	oai:doaj.org-article:fb456fcc79dd48cfb019c88a7a8a1c222021-12-02T20:15:36ZScanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying.1932-620310.1371/journal.pone.0254165https://doaj.org/article/fb456fcc79dd48cfb019c88a7a8a1c222021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0254165https://doaj.org/toc/1932-6203The cellular cortex is an approximately 200-nm-thick actin network that lies just beneath the cell membrane. It is responsible for the mechanical properties of cells, and as such, it is involved in many cellular processes, including cell migration and cellular interactions with the environment. To develop a clear view of this dense structure, high-resolution imaging is essential. As one such technique, electron microscopy, involves complex sample preparation procedures. The final drying of these samples has significant influence on potential artifacts, like cell shrinkage and the formation of artifactual holes in the actin cortex. In this study, we compared the three most used final sample drying procedures: critical-point drying (CPD), CPD with lens tissue (CPD-LT), and hexamethyldisilazane drying. We show that both hexamethyldisilazane and CPD-LT lead to fewer artifactual mesh holes within the actin cortex than CPD. Moreover, CPD-LT leads to significant reduction in cell height compared to hexamethyldisilazane and CPD. We conclude that the final drying procedure should be chosen according to the reduction in cell height, and so CPD-LT, or according to the spatial separation of the single layers of the actin cortex, and so hexamethyldisilazane.Moritz SchuEmmanuel TerriacMarcus KochStephan PaschkeFranziska LautenschlägerDaniel A D FlormannPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 7, p e0254165 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Moritz Schu Emmanuel Terriac Marcus Koch Stephan Paschke Franziska Lautenschläger Daniel A D Flormann Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying.
description	The cellular cortex is an approximately 200-nm-thick actin network that lies just beneath the cell membrane. It is responsible for the mechanical properties of cells, and as such, it is involved in many cellular processes, including cell migration and cellular interactions with the environment. To develop a clear view of this dense structure, high-resolution imaging is essential. As one such technique, electron microscopy, involves complex sample preparation procedures. The final drying of these samples has significant influence on potential artifacts, like cell shrinkage and the formation of artifactual holes in the actin cortex. In this study, we compared the three most used final sample drying procedures: critical-point drying (CPD), CPD with lens tissue (CPD-LT), and hexamethyldisilazane drying. We show that both hexamethyldisilazane and CPD-LT lead to fewer artifactual mesh holes within the actin cortex than CPD. Moreover, CPD-LT leads to significant reduction in cell height compared to hexamethyldisilazane and CPD. We conclude that the final drying procedure should be chosen according to the reduction in cell height, and so CPD-LT, or according to the spatial separation of the single layers of the actin cortex, and so hexamethyldisilazane.
format	article
author	Moritz Schu Emmanuel Terriac Marcus Koch Stephan Paschke Franziska Lautenschläger Daniel A D Flormann
author_facet	Moritz Schu Emmanuel Terriac Marcus Koch Stephan Paschke Franziska Lautenschläger Daniel A D Flormann
author_sort	Moritz Schu
title	Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying.
title_short	Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying.
title_full	Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying.
title_fullStr	Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying.
title_full_unstemmed	Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying.
title_sort	scanning electron microscopy preparation of the cellular actin cortex: a quantitative comparison between critical point drying and hexamethyldisilazane drying.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/fb456fcc79dd48cfb019c88a7a8a1c22
work_keys_str_mv	AT moritzschu scanningelectronmicroscopypreparationofthecellularactincortexaquantitativecomparisonbetweencriticalpointdryingandhexamethyldisilazanedrying AT emmanuelterriac scanningelectronmicroscopypreparationofthecellularactincortexaquantitativecomparisonbetweencriticalpointdryingandhexamethyldisilazanedrying AT marcuskoch scanningelectronmicroscopypreparationofthecellularactincortexaquantitativecomparisonbetweencriticalpointdryingandhexamethyldisilazanedrying AT stephanpaschke scanningelectronmicroscopypreparationofthecellularactincortexaquantitativecomparisonbetweencriticalpointdryingandhexamethyldisilazanedrying AT franziskalautenschlager scanningelectronmicroscopypreparationofthecellularactincortexaquantitativecomparisonbetweencriticalpointdryingandhexamethyldisilazanedrying AT danieladflormann scanningelectronmicroscopypreparationofthecellularactincortexaquantitativecomparisonbetweencriticalpointdryingandhexamethyldisilazanedrying
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Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying.

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