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Review

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

1
Center for Molecular Microscopy, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA
2
Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD 21701, USA
3
National Institute of Allergy and Infectious Diseases, Division of Clinical Research, Integrated Research Facility at Fort Detrick (IRF-Frederick), Frederick, MD 21702, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Cristina Risco
Viruses 2021, 13(4), 611; https://doi.org/10.3390/v13040611
Received: 8 February 2021 / Revised: 18 March 2021 / Accepted: 19 March 2021 / Published: 2 April 2021
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. View Full-Text
Keywords: FIB-SEM; volumeEM; vEM; SARS-CoV-2; virological synapse; segmentation FIB-SEM; volumeEM; vEM; SARS-CoV-2; virological synapse; segmentation
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MDPI and ACS Style

Baena, V.; Conrad, R.; Friday, P.; Fitzgerald, E.; Kim, T.; Bernbaum, J.; Berensmann, H.; Harned, A.; Nagashima, K.; Narayan, K. 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. Viruses 2021, 13, 611. https://doi.org/10.3390/v13040611

AMA Style

Baena V, Conrad R, Friday P, Fitzgerald E, Kim T, Bernbaum J, Berensmann H, Harned A, Nagashima K, Narayan K. 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. Viruses. 2021; 13(4):611. https://doi.org/10.3390/v13040611

Chicago/Turabian Style

Baena, Valentina, Ryan Conrad, Patrick Friday, Ella Fitzgerald, Taeeun Kim, John Bernbaum, Heather Berensmann, Adam Harned, Kunio Nagashima, and Kedar Narayan. 2021. "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" Viruses 13, no. 4: 611. https://doi.org/10.3390/v13040611

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