To begin, launch the software compatible with the in vivo imaging system. Log in and wait for the imaging wizard panel to fully load. Click on the Initialize button on the imaging wizard panel to initiate the cooling of the camera.
Then set the temperature of the heating pad between 30 to 37 degrees Celsius to keep the animals warm. Next, gently pinch the foot pads of the anesthetized animals to assess their pedal reflex. Intraperitoneally inject 10 microliters per gram body weight of sodium luciferin into the shaved animals.
Now align the animal against the camera's center as denoted by the plus symbol in the imaging instrument. Set the imaging time in the image wizard panel to three minutes for luminescence imaging and select the checkboxes for both photo and luminescence. Click on Acquire to take the first measurement.
After all measurements have been recorded, convert the image scale to radiance. Now determine the optimal binning and color scale settings for the images and keep it uniform for all images. Then click on ROI tool on the tool palette panel and use the Place ROI option to select the areas of interest on the image.
Lastly, click on Measure ROIs and copy the resulting data for exporting into the organizing or statistical software of choice. In vivo imaging of the THAI mice showed that areas that lacked fur, such as foot pads, tail, and nose, had higher basal signals. Luciferase signal was influenced by the thyroid hormone status.
Robust signals were observed in the testicular region. The thyroid hormone action was significantly heightened in the brown adipose tissue of cold stressed THAI mice. The treatment did not impact the luciferase signals from the foot pads and tail.
Imaging studies also captured the accumulation and clearance of diclazuril, an endocrine-disrupting compound, over a three-week period.
