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How to prepare Luciferin for in vivo use?
Posted August 5, 2019
Answer
Luciferin is a common reagent used for in vivo imaging of luciferase labeled tumors or other infectious diseases in rats or mice. Below is an example protocol for in vivo bioluminescent imaging using D-Luciferin, free acid.
Materials:
- D-Luciferin, free acid (Cat No. 12501, 12502 and 12503)
- 1X dPBS w/o Ca2+ and Mg2+
- Syringe filter, 0.22 µm
Procedure:
- Prepare a fresh stock solution of D-Luciferin, free acid at 15 mg/mL in 1X dPBS w/o Ca2+ and Mg2+.
- Filter through a 0.22 µm syringe filter to sterilize solution.
- Use immediately, or aliquot into sterilized tubes and freeze at -80°C for up to 6 months.
- Inject the Luciferin solution intra-peritoneally 10-15 minutes before imaging*.
- Inject 10 µL/g of body weight. Each mouse should receive 150 mg Luciferin/kg of body weight.
- Take images every 5-10 minutes for about 40-60 minutes to determine the kinetic curve.
*A kinetic study of luciferin should be performed for each animal model to determine peak signal time.
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