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Related Concept Videos

In-situ Hybridization02:31

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

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Related Experiment Video

Updated: Jun 30, 2026

Combining Multiplex Fluorescence In Situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections
07:36

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

Published on: June 25, 2021

Using Combined Fluorescent In Situ Hybridization With Immunohistochemistry to Co-localize mRNA in Diverse Neuronal

Melanie K Becher1, Kaela Wilson2, Italo Mocchetti1

  • 1Interdisciplinary Program in Neuroscience, Georgetown University Medical Center Washington, DC, USA.

Bio-Protocol
|June 29, 2026
PubMed
Summary

This study presents a novel protocol combining fluorescent in situ hybridization (FISH) and immunohistochemistry (IHC) for sensitive mRNA detection in specific neuronal populations. This method allows for high-resolution, quantitative analysis of gene expression alongside protein localization in the brain.

Keywords:
BrainFluorescent in situ hybridizationImage analysisImmunohistochemistrymRNA

More Related Videos

Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections
09:23

Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections

Published on: March 26, 2016

Related Experiment Videos

Last Updated: Jun 30, 2026

Combining Multiplex Fluorescence In Situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections
07:36

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

Published on: June 25, 2021

Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections
09:23

Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections

Published on: March 26, 2016

Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Understanding gene expression in specific neuronal populations is crucial for brain research.
  • Current methods for mRNA and protein detection have limitations in resolution and quantification.
  • Distinguishing transcriptional changes from protein abundance requires precise detection techniques.

Purpose of the Study:

  • To develop a protocol for sensitive and high-resolution detection of mRNA in single neuronal cell types.
  • To enable simultaneous visualization and quantification of mRNA and protein expression within the same tissue section.
  • To facilitate the study of cellular and molecular diversity in the brain.

Main Methods:

  • Combined fluorescent in situ hybridization (FISH) using ACDBio RNAscope technology with standard immunohistochemistry (IHC).
  • Developed a custom macro code for FIJI/ImageJ to analyze mRNA expression and co-localization.
  • Applied the protocol to murine brain tissue for simultaneous mRNA and protein detection.

Main Results:

  • Achieved sensitive and anatomically resolved detection of mRNA in single neuronal cell types.
  • Enabled visual simultaneous detection of mRNA and protein expression, allowing for co-localization analysis.
  • Demonstrated efficient quantification of distinct mRNA transcripts within specific neuronal subpopulations.

Conclusions:

  • The combined FISH-IHC protocol offers a powerful tool for studying gene expression at the single-cell level.
  • This method enhances the ability to dissect neuronal diversity by correlating mRNA transcripts with protein expression.
  • The protocol is adaptable for detecting various mRNA-protein combinations across different tissue types.