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D-luciferin is a natural substrate of luciferase that catalyzes the production of typical yellow green light by bioluminescent insects.
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In vitro studies have shown that D-luciferin is a natural substrate of the enzyme luciferase (Luc), which catalyzes the production of typical yellow green light in fireflies. This review covers the synthesis of D-luciferin and its derivatives or analogues as substrates or inhibitors of luciferase. The American firefly Photinus pyralis, this enzyme is more commonly used in vitro and optical imaging techniques [1].
In vivo bioluminescence imaging (BLI) using firefly luciferase (Fluc) as the reporter gene and D-luciferin as the substrate is currently the most widely used technology. Plot the total signal intensity relative to the time after D-fluorescein injection to generate a time intensity curve. In addition to the peak signal, signals at fixed time points (5,10,15 and 20 minutes) after D-fluorescein injection were identified as substitutes for the peak signal. The signal in the given time intensity curve is normalized for the peak signal in the curve to represent the time change pattern after D-fluorescein injection [2].
Animal experimental mice [2] were inoculated with HCT116 for 3 days, 5 days, 7 days, 10 days, 12 days, 14 days, 19 days, 21 days, 24 days, and 28 days to observe BLI Luc cells in vivo using a cooled charge coupled device camera system (IVIS imaging system 100). Subcutaneous injection of 100 into mice near the scapula μ 75mg/kg D-fluorescein in L-phosphate buffered saline was placed in an opaque chamber of the imaging system under isoflurane anesthesia. Starting 5 minutes after injection, obtain back luminescent images with an exposure time of 1 second at a rate of one image per minute, until 20 minutes after D-fluorescein injection. Due to the extended time process of light emission, data collection lasts from the 3rd or 5th day to 40 minutes after injection and from the 7th day to 25 minutes. The division is 4, and the field of view is 15 centimeters.