This article describes a method for imaging bacterial infections in anesthetized mice using bioluminescence and X-ray imaging. The technique allows for real-time monitoring of infection progression and localization within the host.
Take an anesthetized mouse pre-infected with a pathogenic bacterial strain engineered to express a genetically integrated bioluminescence operon.
The operon encodes luciferase and a multienzyme complex that synthesizes its substrate.
The luciferase enzyme catalyzes a reaction with these substrates, producing visible luminescence.
Place the mouse in a supine position on the temperature-stabilized platform of an imaging chamber and maintain anesthesia during imaging.
Use the imaging system to capture both bioluminescent and corresponding X-ray images of the mouse.
The bioluminescent image reveals the extent of bacterial infection, while the X-ray image provides an anatomical reference for infection localization.
After imaging, return the mouse to its cage and allow recovery.
Capture bioluminescence images at regular intervals to monitor the real-time progression of the bacterial infection.
Over time, the presence of bioluminescent signals at various locations confirms the spread of the infection throughout the host.
To image the animals, prepare the imaging system by setting the camera parameters and heating the stage. Set the oxygen flow to 1.5 liters per minute and isoflurane to 3.5% and move the mouse to the anesthetic chamber, then to the temperature stabilized stage, following anesthesia. Position the mouse on its back with arms outstretched and fit a nose cone for administration of 2.5% isoflurane during imaging.
Close the door and take bioluminescent images and X-rays of the mouse. When imaging is complete, return the mouse to its cage and monitor it. It should regain consciousness within 3 to 5 minutes.
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