Construction and organization of a BSL-3 cryo-electron microscopy laboratory at UTMB

Michael B. Sherman, Juan Trujillo, Ian Leahy, Dennis Razmus, Robert DeHate, Paul Lorcheim, Mark A. Czarneski, Domenica Zimmerman, Je T.Aime M. Newton, Andrew D. Haddow, Scott C. Weaver

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

A unique cryo-electron microscopy facility has been designed and constructed at the University of Texas Medical Branch (UTMB) to study the three-dimensional organization of viruses and bacteria classified as select agents at biological safety level (BSL)-3, and their interactions with host cells. A 200. keV high-end cryo-electron microscope was installed inside a BSL-3 containment laboratory and standard operating procedures were developed and implemented to ensure its safe and efficient operation. We also developed a new microscope decontamination protocol based on chlorine dioxide gas with a continuous flow system, which allowed us to expand the facility capabilities to study bacterial agents including spore-forming species. The new unified protocol does not require agent-specific treatment in contrast to the previously used heat decontamination. To optimize the use of the cryo-electron microscope and to improve safety conditions, it can be remotely controlled from a room outside of containment, or through a computer network world-wide. Automated data collection is provided by using JADAS (single particle imaging) and SerialEM (tomography). The facility has successfully operated for more than a year without an incident and was certified as a select agent facility by the Centers for Disease Control.

Original languageEnglish (US)
Pages (from-to)223-233
Number of pages11
JournalJournal of Structural Biology
Volume181
Issue number3
DOIs
StatePublished - Mar 1 2013

Keywords

  • Biological safety containment
  • Cryo-electron microscopy
  • Electron tomography
  • Single particle imaging

ASJC Scopus subject areas

  • Structural Biology

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    Sherman, M. B., Trujillo, J., Leahy, I., Razmus, D., DeHate, R., Lorcheim, P., Czarneski, M. A., Zimmerman, D., Newton, J. T. A. M., Haddow, A. D., & Weaver, S. C. (2013). Construction and organization of a BSL-3 cryo-electron microscopy laboratory at UTMB. Journal of Structural Biology, 181(3), 223-233. https://doi.org/10.1016/j.jsb.2012.12.007