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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 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.

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Document Type:Article
Author:Moritz Schu, Emmanuel TerriacORCiD, Marcus Koch, Stephan Paschke, Franziska LautenschlägerORCiD, Daniel A. FlormannORCiD
Parent Title (English):PloS one
First Page:e0254165
Year of first Publication:2021
Release Date:2022/11/18
Tag:actins; artifacts; cultured cells; desiccation; freeze Drying; humans; organosilicon Compounds; scanning electron microscopy; specimen Handling
Impact:03.752 (2021)
Funding Information:Deutsche Forschungsgemeinschaft (10271149) Universität des Saarlandes INM Leibniz Institut für Neue Materialien ulm university/university hospital ulm, ulm, germany: www.uniklinik-ulm.de
Scientific Units:Physical Analytics
Open Access:Open Access
Signature:INM 2021/077
Licence (German):License LogoCreative Commons - CC BY - Namensnennung 4.0 International