1. Anesthetize the rat by placing it in a bell jar filled with methoxyflurane or giving it an i.p. injection of ketamine (100 mL/kg)
2. Pick up the rat and place it in your left hand. Secure its legs behind your fingers and make sure that its neck is extended past your knuckles
3. Hold the rat above the large petri dish of CSF. Cut its head off in a single, firm motion with the large scissors. Place the body neck side down into the body bag
4. Keeping the head submerged in CSF, position it dorsal side up and hold it firmly in place with your left hand. Brace your right hand against the side of the ice bucket and use the scalpel to cut the scalp from end to end along the midline exposing the skull surface
5. Peel the scalp back and hold it away from the skull with your left hand. Insert the angled scissors into the spinal cord hole at the base of the skull. Angling them upward (so as not to touch the brain), cut along the midline of the skull. Then, then make perpendicular cuts along bregma and lambda. Peel the skull back and hold it in place. Make sure you don’t cut into the cortex when you do this!
6. Use the scalpel to remove the cerebellum. Then, slice the brain in half along the midline to separate the hemispheres
7. Tilt the head and free the brain hemispheres from the skull with your scalpel. Use a plastic spoon to transport the brain to the small dish of CSF
8. Spoon some CSF onto the filter paper-covered dish bottom. Transport half the brain to this filter paper. Being careful not to cut through cortex, slice away the remaining brainstem. Make a thin slice along the dorsal surface (this will be your gluing surface). Remove the frontal lobe by making a transverse cut along the blood vessel about 1 cm from the front of the brain
9. To do both hemispheres simulatenously, repeat step 17 for the other half of the brain.
