Cryoprotection of Mouse Brain
Ashley Harms, Asta Zane, Nicole J Corbin-Stein, Gabrielle Childers, Jhodi Webster, Vickie Yang, Woong-Jai Won, Rajesh Gupta
Abstract
This protocol allows for accurate cryoprotection of mouse brain, post-perfusions, to be used for histology. The methods utilize another PFA fixation step followed by sucrose incubation.
Steps
PROCEDURE
Transfer the brain into 5-10ml of 4% PFA solution in PBS for 2 hours at room temperature in a 15 ml conical tube.
Transfer the brain into 30% sucrose solution in PBS, wait until it sinks to the bottom for 48-72 hours at 4°C.
Freeze brain on dry ice and store at minus -80 °C till sectioning.
To freeze the brain cut foil into pieces that will be labeled accordingly. Lay the foil on the top of the dry ice. Put the brain on the foil. Let the brain to freeze. It should change its color to white, and become hard. Wrap the foils edges. Put samples in the box, label the box with your project, and put it to minus -80 °C.
