Cryopreserved Tissue Section Fixation and Staining: A Procedure to Process Frozen Brain Tissue Sections for Laser Microdissection

Submerge the slides in 30-second sequential descending ethanol immersions, followed by crystal violet solution staining for 20 seconds and 5 seconds in 0.5% eosin Y solution.

At the end of the eosin Y incubation, use filter paper to blot the slides dry and dehydrate the slides in sequential ascending ethanol immersions. After the 60-second ethanol immersion, rinse the slides in a container of xylene for 3 minutes.

Next, dry the slides on RNase-free surface at room temperature for 10 seconds before mounting the samples in mounting medium prepared with RNase-free water. After 10 to 20 seconds, transfer the slides onto the microscope microdissection platform.