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

DOI:
10.3791/61709-v

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07:36 min

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June 25, 2021

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, ,
1Department of Pharmacology, School of Medical Sciences,University of New South Wales

Summary

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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 HybridizationFluorescent ImmunohistochemistryFixed Brain SectionsRNAscopeSpatial VisualizationMRNAProteinNeuroscienceCryostatParaformaldehyde FixationDehydrationVibrating MicrotomeCryoprotectantPBS WashHydrophobic BarrierProtease TreatmentRNAscope ProbesAmplificationImmunohistochemistry

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