Cryogenic Electron Microscopy of Rift Valley Fever Virus

Michael B Sherman; Thomas J Smith

doi:10.1007/978-1-0716-4338-9_6

Cryogenic Electron Microscopy of Rift Valley Fever Virus

Methods Mol Biol. 2025:2893:57-72. doi: 10.1007/978-1-0716-4338-9_6.

Authors

Michael B Sherman^{1

2}, Thomas J Smith^{3

4}

Affiliations

¹ Department of Biochemistry and Molecular Biology, University of Texas Medical Branch at Galveston, Galveston, TX, USA. mbsherma@utmb.edu.
² Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, TX, USA. mbsherma@utmb.edu.
³ Department of Biochemistry and Molecular Biology, University of Texas Medical Branch at Galveston, Galveston, TX, USA.
⁴ Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, TX, USA.

PMID: 39671030
DOI: 10.1007/978-1-0716-4338-9_6

Abstract

Rift Valley fever virus (RVFV) is an important livestock and human pathogen. It is also a potential bioweapon owing to its ability to spread by aerosols. It is an enveloped virus containing surface protrusions composed of two viral glycoproteins, G_c and G_n; the viral core contains ribonucleoprotein complexes. We describe the use of cryogenic electron microscopy (Cryo-EM) of single particles to visualize the three-dimensional (3D) organization of RVFV. The spherical RVFV virions have a diameter of 100 nm with icosahedral symmetry (T = 12) and 720 prominent protrusions on their surface.In this chapter we describe the general protocols to prepare virus suspensions for Cryo-EM, image acquisition of virus particles embedded in vitreous water, and their image processing. Because of RVFV being classified as a BSL3 and potentially as a select agent, we also describe the protocol of performing Cryo-EM grid preparation and imaging in our Cryo-EM BSL3 containment, along with the general etiquette of using BSL3 containment for research, including entry into containment, dressing up for work there, manipulation with the agent, grid preparation, transfer of the frozen grids into a microscope, etc., waste disposal, and finally exit from the containment. The microscope decontamination with ClO₂ is briefly described as well because that technique is not widely used in the Cryo-EM field.

Keywords: Biological safety level 3; Cryo-EM; Cryo-EM decontamination; Cryogenic electron microscopy; Image processing; RVFV; Rift Valley fever virus.

MeSH terms

Animals
Cryoelectron Microscopy* / methods
Humans
Image Processing, Computer-Assisted / methods
Imaging, Three-Dimensional / methods
Rift Valley Fever / virology
Rift Valley fever virus* / ultrastructure
Virion / ultrastructure