Cryopreserved Tissue Section Fixation and Staining: A Procedure to Process Frozen Brain Tissue Sections for Laser Microdissection
Published: April 30, 2023
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 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. Prepare the described fixative solutions in clean RNase-free 50 mL tubes. M…
Divulgaciones
The authors have nothing to disclose.
Materials
| 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 |
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Cryopreserved Tissue Section Fixation and Staining: A Procedure to Process Frozen Brain Tissue Sections for Laser Microdissection. J. Vis. Exp. (Pending Publication), e20620, doi: (2023).