Preparing Rhinal Cortex-Hippocampus Slices from a Rat Pup Brain

Take a freshly harvested rat pup brain, dorsal side up, in a culture plate with chilled buffer.

Open the brain hemispheres and remove the tissues covering the hippocampus.

Incise the hemispheres so that they contain the hippocampi and rhinal cortices.

Place both hemispheres with the hippocampi facing up and parallel on a filter paper.

Transfer this assembly to a tissue chopper with the blade perpendicular to the hemispheres.

Cut the hemispheres to the required thickness to obtain slices and transfer them to a culture plate with chilled buffer.

Carefully separate the slices and dissect them to include structurally intact hippocampal and rhinal cortex regions.

Transfer the slices to membranous inserts in a multi-well plate containing media.

Remove the excess buffer from the slices and incubate them for tissue recovery.

Refresh the media. The slices can be used for further analysis.

Transfer the brain, dorsal side up, into a 60-millimeter plate containing 5 milliliters of cold GBSS. Insert fine forceps into the cerebellum, and insert the spatula along the midline to carefully open the hemisphere. Use short, curved forceps to carefully remove the excess tissue that covers the hippocamp