Protocol for using negative pressure isolator systems to study BSL-2 organisms in gnotobiotic murine models

Aidan Pavao; Madeline Graham; Olivia Trofimuk; Mary L Delaney; Vladimir Yeliseyev; Lynn Bry

doi:10.1016/j.xpro.2022.101211

Protocol for using negative pressure isolator systems to study BSL-2 organisms in gnotobiotic murine models

STAR Protoc. 2022 Mar 3;3(1):101211. doi: 10.1016/j.xpro.2022.101211. eCollection 2022 Mar 18.

Authors

Aidan Pavao¹, Madeline Graham¹, Olivia Trofimuk^{1

2}, Mary L Delaney^{1

2}, Vladimir Yeliseyev¹, Lynn Bry^{1

2}

Affiliations

¹ Massachusetts Host-Microbiome Center, Department Pathology, Brigham & Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
² Clinical Microbiology Laboratory, Department of Pathology, Brigham & Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.

Abstract

Here, we present a protocol for the use of negative pressure isolator systems to maintain defined association and contain BSL-2 pathogens in germ-free and gnotobiotic mouse studies. We describe setup and operation of negative pressure isolators with integrated microbiologic procedures, using the BSL-2 pathogen Clostridioides difficile as a working example. This approach supports experimental systems with defined-association mice and enables high-resolution mechanistic studies of pathogen-commensal interactions and their impacts on host phenotypes. For complete details on the use and execution of this protocol, please refer to Girinathan et al. (2021).

Keywords: Microbiology; Microscopy; Model Organisms.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Disease Models, Animal
Germ-Free Life*
Mice
Microbiological Techniques
Symbiosis*

Abstract

Publication types

MeSH terms

Grants and funding