This article describes a method for imaging neuroinflammation in transgenic mice using bioluminescence. The technique involves the use of luciferin to measure astrocytic activation and neuroinflammatory responses.
Begin with an anesthetized transgenic mouse containing pathogenic protein aggregates, which cause neuroinflammation and astrocyte activation, leading to increased luciferase expression.
Remove the fur from its head to prevent light scattering during imaging.
Color the ears with a non-irritating marker to block nonspecific bioluminescence.
Take an appropriate volume of luciferin based on the mouse's body weight and inject it intraperitoneally.
Luciferin enters the bloodstream, reaches the brain, and is taken up by activated astrocytes.
Within the astrocyte, the luciferase oxidizes luciferin, emitting a bioluminescent signal.
Set the imaging parameters and precisely position the mouse on the bioluminescence imaging chamber's heating plate.
Ensure proper anesthesia flow and capture the bioluminescent signal for the desired duration.
Then, select the region of interest and quantify the targeted bioluminescent signal.
The intensity of this signal corresponds to luciferase activity, reflecting astrocytic activation and providing a direct measure of neuroinflammation.
Prior to imaging, shave and depilate the heads of isoflurane-anesthetized mice. Color the ears in black with a non-irritating marker to block unspecific bioluminescence. Weigh the animals prior to injection, then calculate the exact volume of d-luciferin for injection at 150mg/kg of body weight.
Intraperitoneally inject the calculated volume of d-luciferin, and then return each animal to the anesthesia chamber. Start the imaging software. After the imaging system has reached its operating temperature, initialize the system by clicking on the initialize button in the control panel, then select the button's luminescent and photograph.
Set the exposure time to 60 seconds, the binning to medium, the f-stop to 1, and the EM gain to off. Select block and then open, and then under the tab subject height, choose 1.50 from the dropdown menu. 10 minutes after injection with d-luciferin, place the animals onto the heating plate in the imaging chamber, and ensure that their muscles are correctly placed in the anesthesia outlet.
Switch the isoflurane flow from the inhalation chamber to the imaging chamber, and properly close the door of the imaging chamber. Click on the acquire button to measure the bioluminescence, which takes 60 seconds.
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