Cryo-electron microscopy of enveloped viruses using upgraded transmission electron microscope: Influenza type A, B viruses and SARS-CoV-2

Author:

Kordyukova L.V.1ORCID,Moiseenko A.V.2ORCID,Timofeeva T.A.3ORCID,Fedyakina I.T.3ORCID

Affiliation:

1. Belozersky Institite of Physico-Chemucal Biology, Lomonosov Moscow State University

2. Lomonosov Moscow State University

3. D.I. Ivanovsky Institute of Virology, N.F. Gamaleya National Research Center for Epidemiology and Microbiology, Ministry of Health of the Russian Federation

Abstract

Cryo-electron microscopy (cryo-EM) is indispensable for the structural studies of enveloped viruses – dangerous pathogens of humans and animals. Yet, it requires highly specialized equipment as well as careful sample preparation. In this work, the capabilities of transmission electron microscope JEOL JEM-2100 equipped with cryo-transfer holder are used, and preliminary cryo-EM data for influenza A and B virus strains and SARS-CoV-2 inactivated with beta-propiolactone are presented. Image analysis allows: (1) to distinguish “empty” viral particles from “full” ones (containing nucleocapsid); (2) to visualize the lipid bilayer of the viral envelope; (3) identify influenza virus surface antigens and the M1 protein layer combined with the inner lipid monolayer; (4) distinguish different morphology of S-spikes on the surface of inactivated SARS-CoV-2 virions. The developed approach provides good image quality for both fundamental and applied research.

Funder

Russian Foundation for Basic Research

Publisher

Moscow University Press

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