Tracking Bacterial Infection Dynamics in a Mouse Using Bioluminescence

Samenvatting

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 bioluminescence to visualize and compare the in vivo behavior of virulent and attenuated bacterial strains in mice. It outlines the preparation of bioluminescent bacterial suspensions and their intraperitoneal injection into mice. The virulent strain exhibits systemic spread, while the attenuated strain remains localized and is gradually cleared, as indicated by the fading signal.

Protocol

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

1. Inoculation of Animals with Bacterial Strains by Injection
   1. On the morning of injections, remove cryovials of the bacterial strains being tested and thaw at 4 °C for 3-4 h. Thaw 0.5 mL for each mouse that will be injected. Keep vials on ice after thawing and inject mice within 2 h.
   2. After thawing, transfer each cryovial to a new 2 mL tube and centrifuge at 4,500 x g for 10 min, discard the supernatant, and resuspend the cell pellet in 1 mL of 1x PBS (Phosphate-b....

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
0.2 mL tubes with flat caps	ThermoScientific	AB-0620	via Fisher Scientific
1 mL Syringe	BD	22-253-260	via Fisher Scientific
1.5 mL Microcentrifuge Tubes	Fisher Scientific	05-408-129
2.0 mL Cryogenic Vials	Corning	430659	via Fisher Scientific
27G needle	BD	14-821-13B	via Fisher Scientific
Benchtop Centrifuge 5804R	Eppendorf	04-987-372	via Fisher Scientific
Benchtop Microcentrifuge	Sorvall	75-003-287	via Fisher Scientific
IVIS Lumina XRMS Series III In Vivo Imaging System	Perkins and Elmer	CLS136340
Phosphate buffered saline powder	Sigma	P3813-10PAK	Sigma-Aldrich