Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) culture and sample preparation for correlative light electron microscopy

Methods Cell Biol. 2024:187:99-116. doi: 10.1016/bs.mcb.2024.02.031. Epub 2024 Mar 4.

Abstract

Correlative Light Electron Microscopy (CLEM) is a powerful technique to investigate the ultrastructure of specific cells and organelles at sub-cellular resolution. Transmission Electron Microscopy (TEM) is particularly useful to the field of virology, given the small size of the virion, which is below the limit of detection by light microscopy. Furthermore, viral infection results in the rearrangement of host organelles to form spatially defined compartments that facilitate the replication of viruses. With the emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), there has been great interest to study the viral replication complex using CLEM. In this chapter we provide an exemplary workflow describing the safe preparation and processing of cells grown on coverslips and infected with SARS-CoV-2.

Keywords: CLEM; Gridded coverslips; SARS-CoV2; Virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COVID-19* / virology
  • Chlorocebus aethiops
  • Humans
  • Microscopy, Electron / methods
  • Microscopy, Electron, Transmission / methods
  • SARS-CoV-2* / ultrastructure
  • Vero Cells
  • Virus Replication