Parallel Acquisition of Plasma Membrane Ultrastructure and Cytosolic Protein Localisation in Cultured Cells via Correlated Immunogold SEM
| Autor: | Begemann, Isabell, Keller, Ulrike, Nüsse, Harald, Klingauf, Jürgen, Galic, Milos |
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| Přispěvatelé: | Universitäts- und Landesbibliothek Münster |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
610 Medicine and health
Cell Membrane curvature-sensitive protein Reproducibility of Results CLEM immunogold labelling Immunohistochemistry Protein Transport Cytosol scanning electron microscopy SE/BSE BAR domain lcsh:Biology (General) Medicine and health Microscopy Electron Scanning Technical Note Animals Humans Gold ddc:610 lcsh:QH301-705.5 Cells Cultured |
| Zdroj: | Cells, Vol 9, Iss 1329, p 1329 (2020) Cells |
| ISSN: | 2073-4409 |
| Popis: | Scanning electron microscopy (SEM) takes advantage of distinct detectors to visualise secondary and back-scattering electrons. Here, we report an integrated approach that relies on these two detection methods to simultaneously acquire correlated information on plasma membrane topography and curvature-sensitive cytosolic protein localization in intact cell samples. We further provide detailed preparation and staining protocols, as well as a thorough example-based discussion for imaging optimisation. Collectively, the presented method enables rapid and precise analysis of cytosolic proteins adjacent to cellular membranes with a resolution of ~100 nm, without time-consuming preparations or errors induced by sequential visualisation present in fluorescence-based correlative approaches. Finanziert durch den Open-Access-Publikationsfonds der Westfälischen Wilhelms-Universität Münster (WWU Münster). |
| Databáze: | OpenAIRE |
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