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Related Concept Videos

FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

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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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In-situ Hybridization02:31

In-situ Hybridization

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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...
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Related Experiment Video

Updated: May 2, 2026

Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH
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Single-Molecule Fluorescence In Situ Hybridization (smFISH) for RNA Localization Relative to a DNA Locus in Bacteria.

Juan Echeverry1, Yu-Huan Wang1, Sangjin Kim2,3

  • 1Department of Physics, University of Illinois Urbana-Champaign, Urbana, IL, USA.

Methods in Molecular Biology (Clifton, N.J.)
|April 30, 2026
PubMed
Summary

This study presents a single-molecule fluorescence in situ hybridization (smFISH) protocol to visualize RNA and DNA locations within bacterial cells. This method aids in understanding RNA localization relative to gene loci.

Keywords:
BacteriaDNA FISHDNA lociFROSRNA localizationsmFISH

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Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments
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Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments
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Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Cell Biology

Background:

  • Understanding RNA localization within cells is crucial for deciphering gene expression regulation.
  • Subcellular landmarks, such as gene loci (transcription sites), are essential for contextualizing RNA positioning.

Purpose of the Study:

  • To describe a single-molecule fluorescence in situ hybridization (smFISH) protocol for simultaneous labeling of target RNAs and their DNA loci in bacteria.
  • To provide a versatile method for investigating RNA localization in diverse bacterial systems.

Main Methods:

  • Development and description of a single-molecule fluorescence in situ hybridization (smFISH) protocol.
  • Application of smFISH for co-visualization of RNA and DNA in bacterial cells.

Main Results:

  • The described smFISH protocol enables precise localization of RNA molecules relative to their corresponding DNA loci in bacteria.
  • The protocol is demonstrated to be versatile and applicable across different bacterial genes and species.

Conclusions:

  • The smFISH protocol offers valuable insights into bacterial RNA localization dynamics.
  • This technique serves as a powerful tool for studying spatial gene expression in prokaryotic cells.