Frozen Mouse Brain Tissue Sectioning: A Procedure to Obtain Thin Frozen Tissue Sections from Frozen Murine Brain Tissue

To acquire frozen brain tumor tissue sections, first, set the temperature of a cryostat between minus 20 and minus 24 degrees Celsius and place the sample block into to the cryostat chamber for 30 to 60 minutes.

While the sample is equilibrating, clean the chamber and knife holder with 100% ethanol and spray the section brushes with RNase cleaning solution. When the sample is ready, remove the mold and use fresh cutting medium to attach the frozen tissue block to the cryostat specimen disk.

Place the block in the disk holder and align the block with the knife blade. Acquire 10-micrometer sections of the brain using one of the RNase-free brushes to carefully flatten and uncurl the tissue pieces onto the cutting surface.

To mount the brain tissue sections onto slides, smoothly press the positively charged side of a labeled RNase-free PEN glass slide onto the section and place the slide into a storage box inside the chamber. When all the slides have been collected, place the storage box at minus 80 degrees Celsius.