Generation of Tilted Amygdala Slices from a Mouse Brain

Take a mouse brain immersed in an ice-cold buffer.

Remove the cerebellum and trim the anterior section containing the medial prefrontal cortex, or mPFC.

Attach the tissue to an angled agar block, which is glued onto a vibratome specimen stage.

Attach additional agar blocks to provide stability during slicing. Place the stage inside the vibratome cutting chamber filled with oxygenated ice-cold buffer.

Generate tilted slices containing the amygdala that respond to fear stimuli and neuronal projections from the mPFC, which form synaptic connections in the amygdala and regulate its activity.

The tilted slices expose the projections, allowing the study of the functional connectivity between the mPFC and the amygdala.

Cut the slices in half and place them in a chamber containing oxygenated artificial cerebrospinal fluid at room temperature to stabilize the tissue.

Incubate the slices at the physiological temperature to recover the tissue for downstream assays.