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

Abstract

Source: Comba, A. et al. Laser Capture Microdissection of Glioma Subregions for Spatial and Molecular Characterization of Intratumoral Heterogeneity, Oncostreams, and Invasion. J. Vis. Exp. (2020)

This video presents the fixation and staining method to process frozen mouse brain tumor tissue. The processed tissue helps differentiate and obtain a specific population of cells during subsequent laser capture microdissection.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Fixation and Staining of Cryopreserved Brain Tissue Sections

1. To preserve RNA integrity, clean all instruments to be used with RNase cleaning solution. Proceed with the fixation and staining protocol inside a fume hood.
2. Prepare the described fixative solutions in clean RNase-free 50 mL tubes. Make all solutions with RNase-free water on the day of the staining.
3. Same day the laser microdissection will be performed, prepare 100%, 95%,....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Cresyl Violet Acetate	Sigma Aldrich	C5042
Eosin Y	Sigma Aldrich	E4009
RNaseZap RNase Decontamination Solution	Fisher Scientific	AM9780
PEN Membrane Glass Slide (2 µm)	Lieca	1150518
Pinpoint Solution	Zymo Research	D3001-1
Tissue-Plus O.C.T. Compound	Fisher Scientific	23-730-571