MDA-MB-231-Luc and HCT116-Luc cells
Mice: 2 subcutaneous HCT116-Luc tumor-bearing female BALB/c nude mice; 4 naïve female BALB/c nude mice;
Artagain black paper (Strathmore, Catalog# 445-109)
D-Luciferin Firefly, potassium salt（Sciencelight, Catalog# luc001）, for in vivo imaging
Syringe (1ml) and needles (25 x 5/8' gauge)
Anesthesia (Pentobarbital Sodium)
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.)
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.
ImagingPlace mice on black paper in the imaging box and image dorsally to detect tumor cells.
要先注射D-Luciferin Firefly, potassium salt，后麻醉小鼠
在注射D-Luciferin Firefly, potassium salt 后15-35分钟之间检测
小鼠周龄要在4-6周
接种部位要合适