Perfusion of Mouse

Materials

0.9 % Saline solution
10% formalin solution
Avertin
Isofluorane
30 ½ gauge needle
1 mL syringe
Peristaltic pump
Collecting dish
Surgical stage/platform
- Make sure this is level with the edge of the collecting dish to allow for the need to rest during the procedure

Taken from: http://web.jhu.edu/animalcare/rdf/avertin.html
Stock Solution (1.6 g/ml)
- Mix 25 g avertin and 15.5 ml tert-amyl alcohol at room temperture for ~12 hours in a dark bottle.
- Can store stock solutions at room temperature for one year.
Working solution (20 mg/ml)
- Mix 0.5 ml avertin stock solution and 39.5 ml 0.9% saline in dark/foil covered container.
- Filter solution through 0.2 micron filter into a dark/foiled covered container.
- Store solution at 4 degree C.
- Replace working solution each month.
Notes:
- Avertin is light sensitive.
- Degredation products are lethal.
- Always store at 4 degree C
- Do NOT use a solution that is yellow or contains a precipitate.
- Avertin is lipid soluble and may require a larger dose.

Place 30 ½ gauge on end of tubing
Insert one tube into 9% saline solution, and one tube into 10% formalin solution

Image 1: Set up of the two tube system from 0.9% saline solution and 10% formalin solution.

Fill line with 0.9% saline until you have a steady stream of 0.9% saline from the end of the needle.
Turn off peristaltic pump and turn stopcock to allow for formalin solution to flow
Turn on peristaltic pump and pump 10% formalin solution until flow fluid is before the stopcock
- Flow has not entered the main line
Turn off peristaltic pump and turn stopcock to allow for saline to flow
Turn on pump and pump saline through the main line at stated flow rate

Image 3: Overview of the entire perfusion setup (excluding the procedural platform in the collecting dish)