Frozen Mouse Brain Tissue Sectioning: A Procedure to Obtain Thin Frozen Tissue Sections from Frozen Murine Brain Tissue

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)

In this video, we demonstrate the sectioning of a frozen mouse brain tumor tissue using a cryostat. The frozen brain tissue sections so obtained are stored at low temperatures until further analysis.

Protocol

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

1. Sectioning Frozen Brain Tumor Tissues

1. Label 2 µm polyethylene naphthalate (PEN) slides with the sample information. Tissue sections will be placed directly on these slides following sectioning.
2. Set the temperature of the cryostat chamber between -20 to -24 °C. Before sectioning, place the sample block in the cryostat chamber and let it equilibrate to the temperature in the chamber for 30–60 min.
3. Clean the cryostat chamber and the k....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
PEN Membrane Glass Slide (2 µm)	Lieca	1150518
Research Cryostat	Leica	CM3050s
RNaseZap RNase Decontamination Solution	Fisher Scientific	AM9780
Peel Away Disposable Embedding Molds	Electron Microscopy Sciences	70182
Pinpoint Solution	Zymo Research	D3001-1
Tissue-Plus O.C.T. Compound	Fisher Scientific	23-730-571