活体成像小鼠皮下瘤模型实验步骤

1、Luciferin Preparation1. 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.)

2、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.

3、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、ImagingPlace mice on black paper in the imaging box and image dorsally to detect tumor cells.