Bioluminescence Imaging of Acinetobacter baumannii-Infected Burn Wounds in Mice

Abstract

Source: Wang, Y., et al. In Vivo Investigation of Antimicrobial Blue Light Therapy for Multidrug-resistant Acinetobacter baumannii Burn Infections Using Bioluminescence Imaging. J. Vis. Exp. (2017)

This video demonstrates the procedure for imaging burn wound infections in mice inoculated with Acinetobacter baumannii carrying a bioluminescent operon. It demonstrates how luciferase-catalyzed substrate oxidation produces light proportional to bacterial load, enabling real-time monitoring of infection progression using a bioluminescence imaging system.

Protocol

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

1. Bioluminescence Imaging of Infections in Mice

1. Image the mice using a low-light imaging system that includes an intensified charge-coupled device camera, a camera controller, a specimen chamber, and an image processor.
2. Anesthetize the mice with an intraperitoneal injection of a ketamine-xylazine cocktail (100 mg/kg - 20 mg/kg). Lightly touch the palpebral of the mice with a cotton swab; an absence of the palpebral reflex suggests an appropriate anesthetic depth.
3. Start the live imaging software. ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
IVIS	PerkinElmer Inc, Waltham, MA	IVIS Lumina Series III	Pre-clinical in vivo imaging
Light-emitting diode LED	VieLight Inc, Toronto, Canada	415 nm	Light source for illumination
Mouse	Charles River Laboratories, Wilmington, MA	BALB/c	7-8 weeks age, 17-19 g weight
Acinetobacter baumannii	Brooke Army Medical Center, Fort Sam Houston, TX	Clinical isolate	Engineered luminescent strain
Greiner dishes	Sigma-Aldrich Co. LLC	P5112-740EA	35 mm ×10 mm