Cerebellum Dissection and Slice Preparation: A Method to Generate Tissue Slices from Mouse Cerebellum
Cerebellum Dissection and Slice Preparation: A Method to Generate Tissue Slices from Mouse Cerebellum
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In mice, the cerebellum is situated at the base of the forebrain and consists of two lateral hemispheres joined by a midline – vermis. The cerebellum, which contains the majority of the brain's neurons, is vital for neurological studies.
To generate cerebellum slices, begin with a decapitated mouse head. Insert scissors in the foramen magnum – the opening at the skull base. Gently incise the skull and remove it. Next, flip out the brain and cut the optic and trigeminal nerves. Release the brain into a chilled dissection medium with its dorsal side up. Separate the hindbrain from the other brain parts.
Then, cut the underlying cerebellar peduncles – the connection between the cerebellum and brain stem – to detach the cerebellum. Remove the meninges from the cerebellum. Carefully place the cerebellum on a tissue chopper platform perpendicular to the chopper blade. Now, dissect the tissue slices of appropriate thickness. Immediately, wet the slices and transfer them back into the Petri dish containing the dissection medium.
Separate and identify the slices from the vermis. Transfer the selected slices into a well-insert in a culture plate containing a suitable growth medium. Incubate the culture to maintain the structural and functional integrity of the dissected cerebellar slices for further experiments.