title: Construction and organization of a BSL-3 cryo-electron microscopy laboratory at UTMB.
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 200keV 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.
authors.affiliation: Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, TX 77555-1055, USA. mbsherma@utmb.edu
authors.forename: Michael B
authors.initials: MB
authors.lastname: Sherman
authors.forename: Juan
authors.initials: J
authors.lastname: Trujillo
authors.forename: Ian
authors.initials: I
authors.lastname: Leahy
authors.forename: Dennis
authors.initials: D
authors.lastname: Razmus
authors.forename: Robert
authors.initials: R
authors.lastname: Dehate
authors.forename: Paul
authors.initials: P
authors.lastname: Lorcheim
authors.forename: Mark A
authors.initials: MA
authors.lastname: Czarneski
authors.forename: Domenica
authors.initials: D
authors.lastname: Zimmerman
authors.forename: Je T'aime M
authors.initials: JT
authors.lastname: Newton
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Haddow
authors.forename: Scott C
authors.initials: SC
authors.lastname: Weaver
mesh_terms: D020285:Cryoelectron Microscopy; D055032:Electron Microscope Tomography; D017053:Infection Control / Q000458:organization & administration / Q000592:standards; D007753:Laboratories / Q000458:organization & administration* / Q000592:standards