This article describes the procedure for obtaining cryosections of mouse brain tissue. It details the necessary steps for preparing and sectioning frozen brain samples for further analysis.
To obtain cryosections or thin frozen tissue sections of a mouse brain, first, take a frozen block of mouse brain embedded in a suitable embedding medium.
Transfer the frozen block to a cryostat chamber and allow the block’s temperature to equilibrate with the set temperature of the cryostat for subsequent sectioning procedure.
Then, use embedding medium to mount the frozen tissue block to the base of the cryostat specimen disc.
Place the specimen disc into the specimen head of the cryostat.
Additionally, secure a blade in the blade holder of the cryostat.
Using the external handwheel, adjust the position of the specimen head to align the tissue block with the blade.
Once the tissue block and blade are correctly oriented, move the handwheel to initiate sectioning, trimming off the unwanted region of the tissue block that contains excess embedding medium.
As the frozen tissue block advances towards the fixed blade, it gets cut into slices of desired thickness.
Carefully transfer the sliced frozen tissue sections onto positively charged glass slides. The charged glass slides electrostatically attract the frozen tissue sections, facilitating their adhesion to the slide surface.
Finally, store the sections at low temperatures until further analysis.
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.
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