Processing of a Mouse Brain for Downstream Analysis

Take a freshly harvested mouse brain and flash-freeze it in pre-chilled isopentane for rapid tissue preservation.

Next, transfer the brain, cortex side up, into a frozen brain matrix to maintain a cold environment during slicing.

Allow the brain to equilibrate to the matrix temperature. Align it in the matrix along the sagittal and transverse sinuses to ensure symmetrical sections.

Insert chilled blades into the brain, ensuring proper alignment.

Then, press them down evenly to obtain uniform brain sections.

Remove the blades with the sections and position them, rostral side up, on a frozen glass plate.

Organize the sections from rostral to caudal orientation in the correct anatomical order. Now, separate them.

Using a suitable brain reference, locate and excise the region of interest from the section.

Transfer the excised tissue into a pre-chilled vial for downstream analysis.

After removing brains from euthanized adult CD-1 wild-type mice flash freeze the tissue for 60 seconds in either liquid nitrogen or isopentane prechilled with dry ice, and store it at -80 degrees Celsius.

24 hours before dissecting the tissue, place a clean brain matrix on a stack of thawed freezer packs, and