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Technical Note

Parallel Acquisition of Plasma Membrane Ultrastructure and Cytosolic Protein Localisation in Cultured Cells via Correlated Immunogold SEM

1
Institute of Medical Physics and Biophysics, University of Muenster, Robert-Koch-Str. 31, 48149 Münster, Germany
2
Interfaculty Centre ‘Cells in Motion’, University of Muenster, Waldeyerstr. 15, 48149 Münster, Germany
*
Author to whom correspondence should be addressed.
Cells 2020, 9(6), 1329; https://doi.org/10.3390/cells9061329
Received: 1 April 2020 / Revised: 6 May 2020 / Accepted: 14 May 2020 / Published: 26 May 2020
(This article belongs to the Section Cell Motility and Adhesion)
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. View Full-Text
Keywords: scanning electron microscopy; SE/BSE; CLEM; immunogold labelling; curvature-sensitive protein; BAR domain scanning electron microscopy; SE/BSE; CLEM; immunogold labelling; curvature-sensitive protein; BAR domain
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MDPI and ACS Style

Begemann, I.; Keller, U.; Nüsse, H.; Klingauf, J.; Galic, M. Parallel Acquisition of Plasma Membrane Ultrastructure and Cytosolic Protein Localisation in Cultured Cells via Correlated Immunogold SEM. Cells 2020, 9, 1329. https://doi.org/10.3390/cells9061329

AMA Style

Begemann I, Keller U, Nüsse H, Klingauf J, Galic M. Parallel Acquisition of Plasma Membrane Ultrastructure and Cytosolic Protein Localisation in Cultured Cells via Correlated Immunogold SEM. Cells. 2020; 9(6):1329. https://doi.org/10.3390/cells9061329

Chicago/Turabian Style

Begemann, Isabell, Ulrike Keller, Harald Nüsse, Jürgen Klingauf, and Milos Galic. 2020. "Parallel Acquisition of Plasma Membrane Ultrastructure and Cytosolic Protein Localisation in Cultured Cells via Correlated Immunogold SEM" Cells 9, no. 6: 1329. https://doi.org/10.3390/cells9061329

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