To begin, obtain the cerebellum from a 10-day old mouse pup in cold dissecting buffer. After removing the excess buffer, position the cerebellum on the cutting platform in a vertical orientation. Carry out parasagittal slicing at a slice thickness of 350 micrometers.
Gently separate the slices with a fine iris spatula and place them into cold dissection medium. Under the binocular, use two curved iris spatulas to carefully separate the tissue slices from each other. For cultures, use slices derived from the cerebellar vermis situated in the medial corticonuclear zone of the organ.
Using a wide spatula, transfer each tissue slice onto a culture insert. Move each insert carrying a tissue slice into a well in the six-well plate containing the pre-warmed medium. During the incubation, aspirate the used medium with a sterile glass pipette.
Using a five milliliter serological pipette, add one milliliter of fresh basal medium eagle with the pipette's tip leaning against the well's wall without disturbing the tissue.
