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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 7, 2026

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization
11:24

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization

Published on: June 17, 2015

Step-by-step in situ hybridization method for localizing gene expression changes in the brain.

Jorge J Palop1, Erik D Roberson, Inma Cobos

  • 1Department of Neurology, Gladstone Institute of Neurological Disease, University of California, San Francisco, San Francisco, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|October 23, 2010
PubMed
Summary

This study details a comprehensive RNA in situ hybridization protocol for quantitative neuroscience. The method enables precise gene expression analysis in specific brain cells, aiding in understanding development and diseases like Alzheimer's.

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Last Updated: Jun 7, 2026

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization
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Published on: June 17, 2015

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Double Fluorescence in situ Hybridization in Fresh Brain Sections
12:15

Double Fluorescence in situ Hybridization in Fresh Brain Sections

Published on: August 14, 2010

Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Gene expression analysis in the central nervous system is crucial for understanding development, plasticity, aging, and disease.
  • Standard techniques like Western blotting and PCR lack cellular resolution for gene expression patterns.
  • RNA in situ hybridization offers a powerful method for visualizing gene expression within specific cell populations.

Purpose of the Study:

  • To describe a detailed, step-by-step RNA in situ hybridization protocol for adult and embryonic brain sections.
  • To provide a method for quantitative gene expression analysis at the cellular level in neuroscience research.
  • To demonstrate the utility of this technique in studying neuropathogenesis, using Alzheimer's disease as an example.

Main Methods:

  • Detailed protocols for RNase-free preparation, perfusion, fixation, and sectioning of brain tissues.
  • Synthesis of digoxigenin-labeled RNA probes (riboprobes) from cDNA clones.
  • In situ hybridization on floating and mounted sections with nonradioactive immunohistochemical detection for light and fluorescence microscopy, including double labeling.

Main Results:

  • A fully detailed RNA in situ hybridization protocol applicable to adult and embryonic brain sections.
  • Successful nonradioactive detection and double labeling of RNA probes for cellular gene expression analysis.
  • Demonstration of the technique's utility in investigating neuropathogenesis, exemplified by Alzheimer's disease.

Conclusions:

  • RNA in situ hybridization is a highly accessible and powerful technique for exploring gene expression profiles at cellular and regional levels in the brain.
  • This method overcomes limitations of bulk analysis techniques by providing cellular specificity.
  • The described protocol facilitates quantitative neuroscience research and aids in understanding complex neurological conditions.