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

Michael 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 Weaver

Research output: Contribution to journalArticle

4 Citations (Scopus)

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 2013

Fingerprint

Cryoelectron Microscopy
Decontamination
Organizations
Safety
Biological Warfare Agents
Electrons
Centers for Disease Control and Prevention (U.S.)
Spores
Hot Temperature
Gases
Tomography
Viruses
Bacteria

Keywords

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

ASJC Scopus subject areas

  • Structural Biology

Cite this

Construction and organization of a BSL-3 cryo-electron microscopy laboratory at UTMB. / Sherman, Michael; Trujillo, Juan; Leahy, Ian; Razmus, Dennis; DeHate, Robert; Lorcheim, Paul; Czarneski, Mark A.; Zimmerman, Domenica; Newton, Je T Aime M; Haddow, Andrew D.; Weaver, Scott.

In: Journal of Structural Biology, Vol. 181, No. 3, 03.2013, p. 223-233.

Research output: Contribution to journalArticle

Sherman, M, Trujillo, J, Leahy, I, Razmus, D, DeHate, R, Lorcheim, P, Czarneski, MA, Zimmerman, D, Newton, JTAM, Haddow, AD & Weaver, S 2013, 'Construction and organization of a BSL-3 cryo-electron microscopy laboratory at UTMB', Journal of Structural Biology, vol. 181, no. 3, pp. 223-233. https://doi.org/10.1016/j.jsb.2012.12.007
Sherman, Michael ; Trujillo, Juan ; Leahy, Ian ; Razmus, Dennis ; DeHate, Robert ; Lorcheim, Paul ; Czarneski, Mark A. ; Zimmerman, Domenica ; Newton, Je T Aime M ; Haddow, Andrew D. ; Weaver, Scott. / Construction and organization of a BSL-3 cryo-electron microscopy laboratory at UTMB. In: Journal of Structural Biology. 2013 ; Vol. 181, No. 3. pp. 223-233.
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