活体成像皮下成瘤实验操作方法
Materials:
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)
Procedure:
A. 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.)
B. Preparation of tumor cells:
- Trypsinize the cells from flasks, wash once in serum-free medium.
- Resuspend in room temperate serum-free medium to a concentration of 1x106 cells in 100ul. Hold at room temperature until ready for use.
C. 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.
D. Imaging
Place mice on black paper in the imaging box and image dorsally to detect tumor cells.
Materials:
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)
Procedure:
A. 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.)
B. Preparation of tumor cells:
- Trypsinize the cells from flasks, wash once in serum-free medium.
- Resuspend in room temperate serum-free medium to a concentration of 1x106 cells in 100ul. Hold at room temperature until ready for use.
C. 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.
D. Imaging
Place mice on black paper in the imaging box and image dorsally to detect tumor cells.
