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Begin with an anesthetized mouse with experimental autoimmune encephalomyelitis or EAE. This condition causes inflammation in the eyes and the brain.
Place the mouse in an imaging apparatus, aligning the upper back at the center.
Cover the lower portion of the mouse with a black cloth to prevent signal interference from this region.
Use imaging software to record the mouse position.
Take a syringe filled with a reagent containing the bioluminescent substrate and inject it intraperitoneally into the mouse.
The reagent circulates and enters the inflammation site, where the substrate interacts with inflammation-specific enzymes.
The enzyme-substrate reaction produces a bioluminescent signal.
Capture images at regular time intervals to monitor this signal.
The optic nerve lies near the surface, making bioluminescence around the eyes prominent. In contrast, signals from the brain are weak as the skull absorbs the emitted light.
In the image, blue indicates mild inflammation, and red indicates severe inflammation.
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