Correlative confocal and scanning electron microscopy of cultured cells without using dedicated equipment

Javier Casares-Arias; Miguel A Alonso; Álvaro San Paulo; María Ujué González

doi:10.1016/j.xpro.2021.100727

Correlative confocal and scanning electron microscopy of cultured cells without using dedicated equipment

STAR Protoc. 2021 Aug 9;2(3):100727. doi: 10.1016/j.xpro.2021.100727. eCollection 2021 Sep 17.

Authors

Javier Casares-Arias¹, Miguel A Alonso¹, Álvaro San Paulo², María Ujué González²

Affiliations

¹ Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid, 28049 Madrid, Spain.
² Instituto de Micro y Nanotecnología, IMN-CNM, CSIC (CEI UAM+CSIC), Tres Cantos, Madrid 28760, Spain.

Abstract

This protocol enables correlative light and electron microscopy (CLEM) imaging of cell surface features without using dedicated equipment. Cells are cultured and fixed on transparent substrates for confocal microscopy imaging. No conductive coating is employed in the scanning electron microscopy workflow, providing a clean cell surface observation, with fiducial markers assisting alignment of optical and topographical images. This protocol describes CLEM imaging for midbody remnants in MDCK cells but can also be applied to different cell types and surface features. For complete details on the use and execution of this protocol, please refer to Casares-Arias et al. (2020).

Keywords: Cell Biology; Microscopy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
Cells, Cultured / metabolism*
Dogs
Electron Microscope Tomography / methods
Imaging, Three-Dimensional / methods
Madin Darby Canine Kidney Cells
Microscopy, Confocal / methods*
Microscopy, Electron, Scanning / methods*
Microscopy, Fluorescence / methods
Software
Workflow