Preparation of Colliculo-Thalamocortical Slices from a Mouse Pup Brain

Place the brain of a mouse pup on a slide with its dorsal side facing upward.

Trim the front of the brain and reposition it on the flat surface created.

Align the dorsal surface and midline of the brain on the slide. Cut at an angle to align the connected auditory midbrain and forebrain structures. These structures are the inferior colliculus or IC, medial geniculate body or MGB, thalamic reticular nucleus or TRN, and auditory cortex or AC.

Attach the angularly cut brain surface on a vibratome specimen stage with agar blocks, ensuring the hindbrain is propped up on the thinner block.

Transfer the stage into the vibratome and add a buffer to prevent tissue damage.

Commence slicing to locate the IC, MGB, TRN, and AC.

Retrieve the slice containing the connected regions, termed colliculo-thalamocortical slice, and transfer the slice into a holding chamber to preserve tissue viability.

To prepare the cutting stage for slicing, cut a small piece of 3% agar of approximately 1 cubic centimeter to be used as a backstop for the brain, and another piece of 1.5 centimeter by 1.5 millimeter by 3 millimeters to be used as a bump to support the inferior colliculus.

Next, glue the backstop onto th