Using soft X-ray tomography for rapid whole-cell quantitative imaging of SARS-CoV-2-infected cells.

SARS-CoV-2 cells cryo-tomography organelles soft X-ray synchrotron tomography viral replication organelles

Journal

Cell reports methods
ISSN: 2667-2375
Titre abrégé: Cell Rep Methods
Pays: United States
ID NLM: 9918227360606676

Informations de publication

Date de publication:
22 11 2021
Historique:
received: 27 05 2021
revised: 10 09 2021
accepted: 22 10 2021
pubmed: 4 11 2021
medline: 4 11 2021
entrez: 3 11 2021
Statut: ppublish

Résumé

High-resolution and rapid imaging of host cell ultrastructure can generate insights toward viral disease mechanism, for example for a severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. Here, we employ full-rotation soft X-ray tomography (SXT) to examine organelle remodeling induced by SARS-CoV-2 at the whole-cell level with high spatial resolution and throughput. Most of the current SXT systems suffer from a restricted field of view due to use of flat sample supports and artifacts due to missing data. In this approach using cylindrical sample holders, a full-rotation tomogram of human lung epithelial cells is performed in less than 10 min. We demonstrate the potential of SXT imaging by visualizing aggregates of SARS-CoV-2 virions and virus-induced intracellular alterations. This rapid whole-cell imaging approach allows us to visualize the spatiotemporal changes of cellular organelles upon viral infection in a quantitative manner.

Identifiants

pubmed: 34729550
doi: 10.1016/j.crmeth.2021.100117
pii: S2667-2375(21)00181-8
pmc: PMC8552653
doi:

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Pagination

100117

Subventions

Organisme : NIGMS NIH HHS
ID : P30 GM138441
Pays : United States
Organisme : NIGMS NIH HHS
ID : P41 GM103445
Pays : United States

Informations de copyright

© 2021 The Authors.

Déclaration de conflit d'intérêts

The authors declare no competing interests.

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Auteurs

Valentina Loconte (V)

Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Department of Anatomy, University of California, San Francisco, San Francisco, CA, USA.

Jian-Hua Chen (JH)

Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Department of Anatomy, University of California, San Francisco, San Francisco, CA, USA.

Mirko Cortese (M)

Department of Infectious Diseases, Molecular Virology Heidelberg University, Heidelberg, Germany.

Axel Ekman (A)

Department of Anatomy, University of California, San Francisco, San Francisco, CA, USA.

Mark A Le Gros (MA)

Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Department of Anatomy, University of California, San Francisco, San Francisco, CA, USA.

Carolyn Larabell (C)

Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Department of Anatomy, University of California, San Francisco, San Francisco, CA, USA.

Ralf Bartenschlager (R)

Department of Infectious Diseases, Molecular Virology Heidelberg University, Heidelberg, Germany.
German Center for Infection Research, Heidelberg Partner Site, Heidelberg, Germany.
Division Virus-Associated Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Venera Weinhardt (V)

Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany.

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