Related Experiment Video
Updated: May 25, 2026
09:57
Whole Mount RNA Fluorescent in situ Hybridization of Drosophila Embryos
Published on: January 30, 2013
In situ hybridization to cellular RNA
R Zeller1, M Rogers, A G Haramis
1University of Utrecht, The Netherlands.
Current Protocols in Pharmacology
|February 2, 2012
Summary
In situ hybridization visualizes cellular RNA localization in tissues. This method uses radiolabeled probes and autoradiography for precise message detection within complex cell populations.
Area of Science:
- Molecular Biology
- Cell Biology
- Biotechnology
Background:
- Determining the location of specific RNA molecules within cells and tissues is crucial for understanding gene expression and cellular function.
- In situ hybridization (ISH) is a powerful technique for achieving this spatial resolution.
Purpose of the Study:
- To outline the methodology for in situ hybridization to cellular RNA.
- To detail the preparation of tissue samples and the hybridization process for accurate RNA localization.
Main Methods:
- Tissue preparation: Samples can be embedded in paraffin and microtome-sectioned, or frozen and cryostat-sectioned.
- Hybridization: Cellular RNA is hybridized with a specific radiolabeled probe (e.g., (35)S-labeled riboprobes or DNA probes).
- Detection: Hybridized probes are visualized using film or emulsion autoradiography.
Main Results:
- Successful application of in situ hybridization allows for the precise determination of specific RNA messages within complex cellular environments.
- The described methods enable visualization of RNA localization in both paraffin-embedded and frozen tissue sections.
Conclusions:
- In situ hybridization is a versatile and effective technique for mapping RNA distribution in cells and tissues.
- The choice between paraffin or frozen sections depends on experimental needs, with both amenable to probe hybridization and autoradiographic detection.
Related Concept Videos
02:31In-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...
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...
02:07FISH - 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,...
02:57Southern Blot
Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
03:21RNA-seq
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...