For in vivo imaging: The injection amount is determined at 10 μL/g body weight. Each mouse receives 150 mg of D-luciferin potassium salt per kg of body weight (for a mouse weighing 10 g, 100 μL should be injected to provide 1.5 mg of D-luciferin potassium salt).
For in vitro imaging: A working solution of 150 μg/ml is generally used for in vitro cell imaging.
Tips: The injection method, type of animal, and body weight can all affect the emission of the signal. It is necessary to explore these factors appropriately. Once the fluorescent signal reaches the strongest and most stable plateau phase, imaging analysis should be conducted using the corresponding instruments.
The working solution should be ready for use. Ideally, the prepared storage solution can be stored at -20°C for a short period. If necessary, the luciferin solution can be stored at -20°C for up to 3 weeks.
Tips: Storing the above solution at any temperature for an extended period may lead to signal degradation!
They are classified into D-luciferin free acid, D-luciferin potassium salt, and D-luciferin sodium salt. Due to their high water solubility, potassium salt and sodium salt are the two most commonly used detection substrates, with potassium salt being the primary form used for in vivo animal studies.
The kinetics of bioluminescent signals are tissue-dependent. It is recommended to perform imaging on animals every 5-10 minutes, up to a maximum of 40 minutes, to create a kinetic curve for each new model. (Common peak signal time points are around 10-15 minutes, and luminescence from the luciferin reaction generally starts to decay after 30 minutes and completely disappears after 3 hours.)
Luciferin substrates are approximately 280 Da, and both their water solubility and lipid solubility are very good, allowing them to easily penetrate cell membranes and the blood-brain barrier.
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