Generation of Medial Thalamus-Anterior Cingulate Cortex Slices from Mouse Brains

Begin with a severed mouse head. Remove the skin to expose the skull, and open the skull to access the brain.

Remove the olfactory bulbs and sever any nerve connections to extract the brain into ice-cold oxygenated artificial cerebrospinal fluid, or aCSF.

Take the brain, and make two vertical cuts on each half to expose the internal anatomy.

Make two angled cuts to isolate the region containing the medial thalamus-anterior cingulate cortex or MT-ACC pathway. The MT, a part of the thalamus, acts as a relay station via neuronal projections extending to the ACC in the frontal cortex.

Fix the brain on an angled plate using an adhesive, make a cut to unfold the brain and expose the pathway, and then attach it to a vibratome specimen stage.

Place the stage in the vibratome, add a buffer, and generate slices containing the MT-ACC pathway.

Keep the brain slices in a continuous flow of oxygenated aCSF to maintain their viability.

Expose the skull of the animal, and trim off the remaining muscle. Next, peel away the dorsal surface of the skull using rongeurs and trim away the sides. Subsequently, using a spatula, cut the olfactory bulbs and nerve connections along the ventral surface of the