To begin, add one milliliter of basal medium eagle to each well of the six-well plate. And using sterile forceps, place a single cell culture insert into each well. Incubate the plate in a 37 degrees Celsius, 5%carbon dioxide incubator for a minimum of two hours.
After setting up the dissection area within a biological hood, wash the surgical tools in 70%ethanol, followed by PBS. Then, position the euthanized animal in the dissection area and make a small incision with scissors away from the cerebellum to expose the brain. Using tweezers with rounded ends, gently peel off the skull and remove the skull in small pieces to avoid damaging the cerebellar tissue.
Expose the brain tissue to extract the cerebellum and disconnect the three pairs of peduncles using a fine curved iris spatula to separate the cerebellar cortex from the brain stem. Then, slide the spatula between the cerebellum, the superior colliculus, and the brain stem to retrieve the cerebellum.
