Laser Capture Microdissection to Isolate Purkinje Cells from a Human Post-Mortem Cerebellum Tissue

Take a human postmortem cerebellar tissue containing the Purkinje cell, granule, and molecular cell layers.

Place it on a cryostat sample holder containing partially frozen embedding media.

Allow the media to freeze completely, securing the tissue.

Transfer this assembly into the cryostat cutting arm and allow the tissue to acclimatize to the cryostat temperature.

Move the tissue closer to the blade and trim it to expose the cortex layers.

Place an anti-roll plate above the blade and obtain cerebellar sections.

Transfer these sections to a membrane slide.

Incubate in increasing ethanol concentrations for tissue fixation and dehydration.

Then, incubate in xylene.

Dry the slide and place it on a UV laser capture microscope stage.

Position the collection cap over the cerebellar tissue.

Microscopically, focus on the Purkinje cells.

Using the laser, capture and collect the Purkinje cells for further analysis.

First, cut a small piece of tissue for sectioning, making sure that the section is approximately 95% cerebellar cortex. Return the remaining tissue either to dry ice or to a freezer at minus 80 degrees Celsius. Next, begin slowly adding OCT on top of the chuck with a slow circular