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

Cells. 2020 May 26;9(6):1329. doi: 10.3390/cells9061329.


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.

Keywords: BAR domain; CLEM; SE/BSE; curvature-sensitive protein; immunogold labelling; scanning electron microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Membrane / ultrastructure*
  • Cells, Cultured
  • Cytosol / metabolism*
  • Gold / chemistry
  • Humans
  • Immunohistochemistry
  • Microscopy, Electron, Scanning*
  • Protein Transport
  • Reproducibility of Results


  • Gold