Preparation of Brain Sections Using a NMDG-Based Protective Recovery Method

Begin with an isolated brain. Quickly submerge the brain in a pre-chilled artificial cerebrospinal fluid or aCSF to pause the metabolic processes.

The aCSF contains NMDG, a large organic cation that reduces neuronal cell injury and death.

The aCSF also contains HEPES, which maintains pH and salt concentration and prevents cell swelling and damage.

Transfer the brain onto a Petri dish containing a filter paper.

Trim the brain to obtain the desired region.

Fix the brain block in the specimen holder using a suitable glue and embed it in the agarose.

Position the specimen holder in the slicer.

Fill the slicer's reservoir with the pre-chilled aCSF and aerate the solution to ensure an optimum oxygen supply.

Obtain sections of the brain block.

Transfer the sections into the prewarmed aCSF to restart the metabolic processes, enabling the sections to recover.