Cryopreservation of Murine Brain by Snap Freezing: A Technique to Rapidly Freeze Whole Mouse Brain Specimen for Histological Analysis

To prepare for the cryopreservation, fill a jar with cold isopentane/2-methylbutane and place the jar into a container filled with liquid nitrogen. While the solvent is chilling, blot the brain dry on a piece of filter paper and place the brain into a labeled cryomold.

Carefully add approximately 5 milliliters of optimal temperature cutting medium to the center of the cryomold and place the brain into the mold in the desired orientation. Using clean forceps, quickly place the cryomold into the chilled isopentane/2-methylbutane for 30 to 40 seconds.

Once the cutting medium has solidified, transfer the cryomold onto dry ice and wrap the cryomold and snap-frozen brain tissue in aluminum foil for minus 80 degrees Celsius storage.