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JoVE Journal Neuroscience
Combining Multiplex Fluorescence In Situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections
 

Combining Multiplex Fluorescence In Situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections

Article DOI: 10.3791/61709-v • 07:36 min • June 25th, 2021
June 25th, 2021 •
1, 1, 1
1Department of Pharmacology, School of Medical Sciences, University of New South Wales

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This protocol describes a method for combining fluorescence in situ hybridization (FISH) and fluorescence immunohistochemistry (IHC) in both fresh frozen and fixed mouse brain sections, with the goal of achieving multilabel FISH and fluorescence IHC signal. IHC targeted cytoplasmic and membrane attached proteins.

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Keywords: Multiplex Fluorescence In Situ Hybridization Fluorescent Immunohistochemistry Fresh Frozen Tissue Fixed Brain Sections RNAscope Spatial Visualization MRNA Protein Neuroscience Tissue Sectioning Cryostat Paraformaldehyde Fixation Dehydration Vibrating Microtome Cryoprotectant PBS Wash Hydrophobic Barrier Protease Treatment RNAscope Probes Amplification Immunohistochemistry
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