Diagnostic Electron Microscopy of Viruses With Low-voltage Electron Microscopes

J Histochem Cytochem. 2020 Jun;68(6):389-402. doi: 10.1369/0022155420929438. Epub 2020 May 21.

Abstract

Diagnostic electron microscopy is a useful technique for the identification of viruses associated with human, animal, or plant diseases. The size of virus structures requires a high optical resolution (i.e., about 1 nm), which, for a long time, was only provided by transmission electron microscopes operated at 60 kV and above. During the last decade, low-voltage electron microscopy has been improved and potentially provides an alternative to the use of high-voltage electron microscopy for diagnostic electron microscopy of viruses. Therefore, we have compared the imaging capabilities of three low-voltage electron microscopes, a scanning electron microscope equipped with a scanning transmission detector and two low-voltage transmission electron microscopes, operated at 25 kV, with the imaging capabilities of a high-voltage transmission electron microscope using different viruses in samples prepared by negative staining and ultrathin sectioning. All of the microscopes provided sufficient optical resolution for a recognition of the viruses tested. In ultrathin sections, ultrastructural details of virus genesis could be revealed. Speed of imaging was fast enough to allow rapid screening of diagnostic samples at a reasonable throughput. In summary, the results suggest that low-voltage microscopes are a suitable alternative to high-voltage transmission electron microscopes for diagnostic electron microscopy of viruses.

Keywords: STEM; infectious disease; negative staining; scanning transmission electron microscopy; thin sectioning; transmission electron microscopy; ultrastructure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Hep G2 Cells
  • Humans
  • Microscopy, Electron / instrumentation
  • Microscopy, Electron / methods*
  • Staining and Labeling
  • Viruses / isolation & purification
  • Viruses / metabolism
  • Viruses / ultrastructure*