JoVE Encyclopedia of Experiments
Microbiology
0 views • 2:40 min • March 31st, 2026
Begin with an anesthetized mouse with a Petri dish lid placed under its abdomen to keep the back horizontal.
The mouse exhibits a burn wound on its back that has been infected with Acinetobacter baumannii, a Gram-negative bacterial pathogen.
The bacteria are genetically engineered to carry a bioluminescence operon.
Position the mouse inside a bioluminescence imaging chamber maintained at an optimum temperature and begin imaging.
At the infection site, the luxA and luxB genes of the bioluminescence operon encode the luciferase enzyme. The luxCDE genes encode enzymes that synthesize the luciferase substrate.
Luciferase catalyzes substrate oxidation and produces visible light, making the bacteria luminescent.
The bioluminescence signal indicates the presence of metabolically active bacteria at the wound site.
Using imaging software, quantify the bioluminescence signal intensity to estimate the viable bacterial load.
Immediately following the inoculation, carry out bioluminescence imaging by starting the live imaging software. In the control panel, click Initialize. Then wait until the color of the temperature box turns green, indicating that the temperature of the stage in the specimen chambe
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