JoVE Encyclopedia of Experiments
Neuroscience
0 views • 3:59 min • July 8th, 2025
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.
Gently scoop out the intact brain into the beaker of prechilled NMDG-HEPES aCSF and allow the brain to uniformly cool for about 1 minute. Now, use a large spatula to transfer the brain from the beaker onto the Petri dish, covered with filter paper.
Trim and mount the brain, according to the preferred angle of slicing and desired brain
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