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Cited by 10Advanced Electron Microscopy Techniques and ApplicationsEnzyme Structure and FunctionAdvanced Materials Characterization Techniques

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UVC inactivation of pathogenic samples suitable for cryo-EM analysis

Open access
Depelteau, Jamie S., Renault, Ludovic, Althof, Nynke, Cassidy, C. Keith, Mendonça, Luiza M., Jensen, Grant J., Resch, Guenter P., Briegel, Ariane·2022·Communications Biology
DOI 10.1038/s42003-021-02962-wPMC PMC8752862

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Abstract

Cryo-electron microscopy has become an essential tool to understand structure and function of biological samples. Especially for pathogens, such as disease-causing bacteria and viruses, insights gained by cryo-EM can aid in developing cures. However, due to the biosafety restrictions of pathogens, samples are often treated by chemical fixation to render the pathogen inert, affecting the ultrastructure of the sample. Alternatively, researchers use in vitro or ex vivo models, which are non-pathogenic but lack the complexity of the pathogen of interest. Here we show that ultraviolet-C (UVC) radiation applied at cryogenic temperatures can be used to eliminate or dramatically reduce the infectivity of Vibrio cholerae and the bacterial virus, the ICP1 bacteriophage. We show no discernable structural impact of this treatment of either sample using two cryo-EM methods: cryo-electron tomography followed by sub-tomogram averaging, and single particle analysis (SPA). Additionally, we applied the UVC irradiation to the protein apoferritin (ApoF), which is a widely used test sample for high-resolution SPA studies. The UVC-treated ApoF sample resulted in a 2.1 Å structure indistinguishable from an untreated published map. This research demonstrates that UVC treatment is an effective and inexpensive addition to the cryo-EM sample preparation toolbox.

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Metadata

Authors
Depelteau, J. S., Renault, L., Althof, N., Cassidy, C. K., Mendonça, L. M., Jensen, G. J., Resch, G. P., & Briegel, A.
Affiliations
  • Leiden University (NL)
  • University of Oxford (GB)
  • California Institute of Technology (US)
  • Brigham Young University (US)
Corresponding author
Briegel, Ariane
ORCID
Venue
Communications Biology
Publisher
Springer Science and Business Media LLC
Published
2022-01-11
Volume
5
Issue
1
Pages
29-29
ISSN
2399-3642 (ISSN-L)
DOI
10.1038/s42003-021-02962-w
PMC
PMC8752862
OpenAlex
W4220769548
Cited by
10
References
4
Citations by year
2026: 1 · 2025: 2 · 2024: 1 · 2023: 4 · 2022: 2
FWCI
1.12
Type
journal article
Language
en
Open access
Licence
Locations
  • Communications Biology (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Leiden Repository (Leiden University) (submittedVersion, cc-by)
  • CaltechAUTHORS (California Institute of Technology) (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Utrecht University Repository (Utrecht University) (submittedVersion, other-oa)
  • Data Archiving and Networked Services (DANS) (submittedVersion, cc-by)
Funders
  • Nederlandse Organisatie voor Wetenschappelijk Onderzoek (737.016.004)
  • EC | Horizon 2020 Framework Programme (Instruct-ULTRA ID: 731005)
  • RCUK | Biotechnology and Biological Sciences Research Council (BB/S003339/1)
  • U.S. Department of Health & Human Services | NIH | Center for Information Technology (AI150464)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Enzyme Structure and Function
  • Physical Sciences › Materials Science › General Materials Science › Advanced Materials Characterization Techniques
Keywords
cryo-EM sample preparation, pathogen inactivation, Vibrio cholerae, apoferritin, cryo-electron tomography, subtomogram averaging, single-particle analysis
MeSH terms
Bacteria, Chemotaxis, Ultraviolet Rays, Vibrio cholerae, Viruses, Cryoelectron Microscopy

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