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*Avertin
*Isofluorane
*30 ½ gauge needle
*1 mL syringe
*Peristaltic pump
*Notes:
**Avertin is light sensitive.
**Degredation Degradation 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 dosein an obese animal.
==Perfusion Pump Set Up==
*Place 30 ½ gauge needle on end of tubing*Insert one tube into 0.9% 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.''
[[File:perfusionbottle.png]]
==Animal Procedure==
*Place mouse in isofluorane chamber and wait for slowing of heart beat**Approximately 1 heartbeat/secondlightly anesthetize the animal.
*Remove mouse from chamber
*For mouse anesthesia, administer 0.8 ml/20 g (of mouse body weight) Avertin through intraperitoneal injection with 30 ½ gauge needle.
*Cut open the diaphragm using standard scissors.
**Be careful to not pierce the heart.
*Using clamp scissors, grab at the base of the sterum sternum, cut through the ribcage and lift to exposure expose the heart.
*Transfer the mouse to the procedural stage/platform.
*Insert the 30 ½ gauge needle (from the tubing with saline/10% formalin solution) into the apex of the left ventricle, being careful to keep the tip of the needle in the lumen of the ventricle.*Immediately after inserting the needle into the left ventricle, cut the aorta right ventricle using standard scissors.
**This allows for the blood to flow out of the mouse and drain into collecting dish.
*Perfuse with saline solution for 10 minutes.
*With the pump still on, remove needle from left ventricle.
**Mouse is now fixed for tissue collection.
*When collecting tissues, keep tissues in 10% formalin solution for ~24 hours after collecting and place in at 4 degree C.
*Transfer tissues to PBS after for long-term storage at 4 degree C.
