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Title: | Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying |
Authors: | Schu, Moritz; Terriac, Emmanuel; Koch, Marcus; Paschke, Stephan; Lautenschläger, Franziska; Flormann, Daniel A.D. |
Publishers version: | https://doi.org/10.1371/journal.pone.0254165 |
URI: | https://oa.tib.eu/renate/handle/123456789/8214 https://doi.org/10.34657/7252 |
Issue Date: | 2021 |
Published in: | PLOS ONE 16 (2021), Nr. 7 |
Journal: | PLOS ONE |
Volume: | 16 |
Issue: | 7 |
Page Start: | e0254165 |
Publisher: | San Francisco, California, US : PLOS |
Abstract: | 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. |
Keywords: | actin; hexamethyldisilazane; organosilicon derivative; unclassified drug; actin; hexamethylsilazane; organosilicon derivative; Article; cell dehydration; cell fixation; cell membrane; cell permeabilization; cell size; cell structure; cellular cortex; chemical procedures; comparative study; concentration (parameter); critical point drying; incubation time; procedures concerning cells; quantitative analysis; scanning electron microscopy; structure analysis; artifact; cell culture; chemistry; desiccation; freeze drying; human; procedures; scanning electron microscopy; specimen handling; Actins; Artifacts; Cells, Cultured; Desiccation; Freeze Drying; Humans; Microscopy, Electron, Scanning; Organosilicon Compounds; Specimen Handling |
Type: | article; Text |
Publishing status: | publishedVersion |
DDC: | 500 610 |
License: | CC BY 4.0 Unported |
Link to license: | https://creativecommons.org/licenses/by/4.0/ |
Appears in Collections: | Medizin |
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Schu, Moritz, Emmanuel Terriac, Marcus Koch, Stephan Paschke, Franziska Lautenschläger and Daniel A.D. Flormann, 2021. Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying. 2021. San Francisco, California, US : PLOS
Schu, M., Terriac, E., Koch, M., Paschke, S., Lautenschläger, F. and Flormann, D. A. D. (2021) “Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying.” San Francisco, California, US : PLOS. doi: https://doi.org/10.1371/journal.pone.0254165.
Schu M, Terriac E, Koch M, Paschke S, Lautenschläger F, Flormann D A D. Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying. Vol. 16. San Francisco, California, US : PLOS; 2021.
Schu, M., Terriac, E., Koch, M., Paschke, S., Lautenschläger, F., & Flormann, D. A. D. (2021). Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying (Version publishedVersion, Vol. 16). Version publishedVersion, Vol. 16. San Francisco, California, US : PLOS. https://doi.org/https://doi.org/10.1371/journal.pone.0254165
Schu M, Terriac E, Koch M, Paschke S, Lautenschläger F, Flormann D A D. Scanning electron microscopy preparation of the cellular actin cortex: A quantitative comparison between critical point drying and hexamethyldisilazane drying. 2021;16(7). doi:https://doi.org/10.1371/journal.pone.0254165
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