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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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Updated: Jun 4, 2026

In Situ Hybridization for the Precise Localization of Transcripts in Plants
12:15

In Situ Hybridization for the Precise Localization of Transcripts in Plants

Published on: November 23, 2011

In situ hybridization.

S R Runge1, Z Hu, M S Runge

  • 1Division of Cardiology, University of Texas Medical Branch, Galveston, TX.

Methods in Molecular Medicine
|February 19, 2011
PubMed
Summary
This summary is machine-generated.

In situ hybridization (ISH) detects mRNA in cells and tissues using radioactive probes for reliable results. This technique requires careful execution of multiple steps for accurate visualization of cellular targets.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • In situ hybridization (ISH) is a key technique for detecting messenger RNA (mRNA) within cellular and tissue contexts.
  • The method involves synthesizing a nucleic acid probe with a sequence complementary to the target mRNA.
  • Visualization of the target can be achieved through autoradiography when using radioactive probes.

Purpose of the Study:

  • To describe the methodology of in situ hybridization (ISH).
  • To compare the efficacy of radioactive versus nonradioactive probes in ISH.
  • To emphasize the importance of precision in performing ISH.

Main Methods:

  • Synthesis of a sequence-complementary nucleic acid probe.
  • Incorporation of radioactive nucleotides into the probe for detection.
  • Utilization of autoradiography for visualizing the hybridized probe-target complex.
  • Comparison with nonradioactive (digoxigenin-based) probes.

Main Results:

  • Radioactive probes demonstrated more reliable and accurate results compared to nonradioactive probes in our experience.
  • The ISH technique, while not inherently difficult, demands meticulous attention to detail throughout its multiple steps.
  • Successful application of ISH is contingent upon careful and precise execution.

Conclusions:

  • Radioactive probes are recommended for in situ hybridization due to superior reliability and accuracy.
  • Careful and precise execution of all procedural steps is critical for obtaining meaningful results in ISH.
  • In situ hybridization remains a valuable tool for mRNA detection in biological research.