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

Cell Rep Methods. 2021 Nov 22;1(7):100117. doi: 10.1016/j.crmeth.2021.100117. Epub 2021 Oct 28.

Abstract

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.

Keywords: SARS-CoV-2; cells; cryo-tomography; organelles; soft X-ray; synchrotron; tomography; viral replication organelles.

Publication types

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

MeSH terms

  • COVID-19* / diagnostic imaging
  • Epithelial Cells
  • Humans
  • Imaging, Three-Dimensional / methods
  • SARS-CoV-2*
  • Tomography, X-Ray / methods