活體成像皮下成瘤實驗操作方法

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:

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.

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:

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.

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.