Perfusion of Mouse

From Bridges Lab Protocols
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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

Surgical tools:

  • Dissecting scissors
  • Forceps
  • Clamp scissors
  • Standard scissors

Avertin Preparation

  • 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.