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- Title
FIB-SEM as a Volume Electron Microscopy Approach to Study Cellular Architectures in SARS-CoV-2 and Other Viral Infections: A Practical Primer for a Virologist.
- Authors
Baena, Valentina; Conrad, Ryan; Friday, Patrick; Fitzgerald, Ella; Kim, Taeeun; Bernbaum, John; Berensmann, Heather; Harned, Adam; Nagashima, Kunio; Narayan, Kedar; Risco, Cristina
- Abstract
The visualization of cellular ultrastructure over a wide range of volumes is becoming possible by increasingly powerful techniques grouped under the rubric "volume electron microscopy" or volume EM (vEM). Focused ion beam scanning electron microscopy (FIB-SEM) occupies a "Goldilocks zone" in vEM: iterative and automated cycles of milling and imaging allow the interrogation of microns-thick specimens in 3-D at resolutions of tens of nanometers or less. This bestows on FIB-SEM the unique ability to aid the accurate and precise study of architectures of virus-cell interactions. Here we give the virologist or cell biologist a primer on FIB-SEM imaging in the context of vEM and discuss practical aspects of a room temperature FIB-SEM experiment. In an in vitro study of SARS-CoV-2 infection, we show that accurate quantitation of viral densities and surface curvatures enabled by FIB-SEM imaging reveals SARS-CoV-2 viruses preferentially located at areas of plasma membrane that have positive mean curvatures.
- Subjects
VIRUS diseases; SARS-CoV-2; FOCUSED ion beams; SCANNING electron microscopy; ELECTRON beams; ELECTRON microscopy
- Publication
Viruses (1999-4915), 2021, Vol 13, Issue 4, p611
- ISSN
1999-4915
- Publication type
Article
- DOI
10.3390/v13040611