活体成像小鼠皮下瘤模型实验步骤
Luciferin Preparation
1. Prepare a stock solution of luciferin at 15mg/ml in DPBS. Filter sterilize through a 0.2 um filter.
2. Prepare enough to inject 10 ul/g of body weight. Each mouse should receive 150 mg luciferin/kg body weight. (e.g. For a 20 g mouse, inject 200 ul to deliver 3 mg of luciferin.)
Preparation of tumor cells:
1. Trypsinize the cells from flasks, wash once in serum-free medium.
2. Resuspend in room temperate serum-free medium to a concentration of 1x106 cells in 100ul. Hold at room temperature until ready for use.
Injections:
1. Inject mice with firefly luciferin (150 mg/kg) by intraperitoneal injection using a 25 x 5/8” gauge needle.
2. After 7-8 minutes, anesthetize mice by intraperitoneal injection of pentobarbital sodium using a 25 x 5/8” gauge needle.
3. (1) Inject tumor cells (1x106 cells in 100ul) subcutaneously into the dorsal flank of the mouse.
(2) For the two HCT116-Luc tumor-bearing mice, no cells were inoculation to the mice.
4
Imaging
Place mice on black paper in the imaging box and image dorsally to detect tumor cells.
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