Real-Time Imaging of Bacterial Infection in a Mouse Model Using Bioluminescence

Abstract

Source: Valentine, M. E., et al., Generation of In-Frame Gene Deletion Mutants in Pseudomonas aeruginosa and Testing for Virulence Attenuation in a Simple Mouse Model of Infection. J. Vis. Exp. (2020)

This video demonstrates the use of a bioluminescence-based imaging approach to monitor bacterial infection in a live mouse model. Sequential bioluminescent and X-ray imaging enables real-time visualization and anatomical localization of infection spread throughout the host.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

Visualization of the Infection with Bioluminescence

1. To visualize the progress of the infection, insert a chromosomal bioluminescent operon (luxCDABE) into the Pseudomonas aeruginosa PGN5 and VE2 strains tested. The plasmids and protocol used to label these strains were developed in the Schweizer lab and may not be compatible with all species/strains of bacteria. Importantly, visualization of the infection is optional; thus, genomic insertion of this operon is not necessary to perform the mortality s....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Accu block Digital Dry Bath	Labnet	NC0205808	via Fisher Scientific
Cabinet Incubator	VWR	1540
Isothesia Isoflurane	Henry Schein Animal Health	29405
IVIS Lumina XRMS Series III In Vivo Imaging System	Perkins and Elmer	CLS136340
MicroPulser Electroporator	BioRad	1652100
Prism 7	GraphPad		https://www.graphpad.com/scientific-software/prism/
SmartSpec Plus Spectrophotometer	Bio-Rad	170-2525	or preferred method/vendor
XAF-8 Anesthesia System Filters	Perkins and Elmer	118999
XGI 8 Gas Anesthesia System	Caliper Life Sciences/Xenogen