This article describes a method for preparing coronal brain slices from mouse brains, focusing on the amygdala and its connections with the medial prefrontal cortex (mPFC). The procedure involves careful dissection and slicing to maintain neuronal viability for further study.
Take a mouse brain in an ice-cold buffer and remove its posterior portion containing the cerebellum and the anterior portion with the medial prefrontal cortex, or mPFC.
The middle portion contains amygdala regions that process emotions like fear and anxiety.
Move the brain section onto a filter paper to remove the excess buffer.
Glue it onto a stage and fix an agar block behind the section to prevent its movement during slicing.
Place this stage into a vibratome cutting chamber with the ice-cold buffer.
Cut the section to obtain a thin coronal brain slice.
This exposes the amygdala regions, which contain synaptic connections between the amygdala neurons and the projections of mPFC neurons that regulate the amygdala's responses.
Cut the slice into two halves, each containing an amygdala region.
Next, transfer them to an interface chamber containing oxygenated artificial cerebrospinal fluid.
Recover the slices at physiological temperature to maintain neuronal viability.
After obtaining the brain, use a scalpel to remove the cerebellum and interior part of the brain, and then cool the middle portion of the brain in ice-cold cutting solution as before. Blot the brain with filter paper, and then glue it to the vibratome stage.
Again, fix an additional agar block behind the brain for stability while slicing. After fitting the stage into the cutting chamber filled with 4 degrees Celsius cutting solution, cut 320-micron coronal sections through the amygdala. Cut them in half and collect the sections in the interface chamber as before.
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