In Situ Hybridization
In Situ Hybridization - Related Videos
08:00
Whole-Mount In Situ Hybridization
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2023
Whole-mount in situ hybridization (WMISH) is a common technique used for visualizing the location of expressed RNAs in embryos. In this process, synthetically produced RNA probes are first complementarily bound, or "hybridized," to the transcripts of target genes. Immunohistochemistry or fluorescence is then used to detect these RNA hybrids, revealing spatial and temporal patterns of gene expression. Unlike traditional in situ hybridization techniques, which require thin tissue sections whose...
12:15
In Situ Hybridization for the Precise Localization of Transcripts in Plants
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Cited by 68 •
2011
The in situ hybridization protocol described here allows a direct localization of mRNA and small RNA expression at the cellular level with high sensitivity and specificity. The procedure is optimized for paraffin-embedded plant tissue sections, is applicable to a wide range of plants and tissues, and can be completed within ten days.
15:42
Double Whole Mount in situ Hybridization of Early Chick Embryos
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Cited by 4 •
2008
This video demonstrates 2-color whole mount in situ hybridization, a method by which the spatial and temporal expression pattern of 2 different genes can be visualized in young chick embryos. This method was originally introduced by David Wilkinson, Domingos Henrique, Phil Ingham and David Ish -Horowicz.
02:31
In-situ Hybridization
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2021
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...
12:21
MicroRNA In situ Hybridization for Formalin Fixed Kidney Tissues
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Cited by 11 •
2013
This article describes an in situ hybridization protocol optimized for colormetric detection of microRNA expression in formalin fixed kidney sections.
FAQs
Specificity comes from base-pairing between the labeled probe and its matching DNA or RNA sequence. A probe is designed to recognize a particular target, allowing the detected signal to be associated with that sequence rather than with genetic material throughout the specimen. This molecular matching supports precise localization of genes, messenger RNA, chromosome regions, or pathogen sequences.
The probe provides both recognition and detectability. Its complementary sequence identifies the target, while the attached fluorescent, enzymatic, or other detectable tag converts that binding event into an observable signal. The selected label therefore affects how researchers visualize the target and connect its molecular location with a specific cell, tissue region, or chromosome.
Preservation maintains the spatial arrangement in which the target sequence occurs. Instead of measuring only whether a DNA or RNA sequence is present, researchers can relate the signal to particular cells, tissue regions, or whole-organism locations. That spatial information helps connect genetic information with tissue organization, developmental patterns, cell identity, and disease-associated changes.
Fluorescence in situ hybridization, or FISH, is especially associated with cytogenetics and genome analysis, where fluorescent signals can reveal chromosome regions. RNA in situ hybridization focuses on locating RNA, including messenger RNA, within biological specimens. The distinction reflects different research questions: chromosome and genome organization versus gene expression, cell identity, development, or tissue patterning.
A typical workflow begins with a preserved cell, tissue, or whole-organism specimen, followed by exposure to a labeled probe designed for the sequence of interest. After the probe binds its complementary target, the attached tag is visualized through fluorescence, an enzymatic reaction, or another detection approach. The resulting signal is interpreted according to its spatial distribution.
This approach is valuable when location is central to the biological question. It can show where genes are expressed, where messenger RNA accumulates, which chromosome regions contain a sequence, or where pathogen material is distributed. Such information adds cellular and tissue context that a measurement lacking spatial resolution would not provide on its own.
RNA in situ hybridization can reveal the distribution of messenger RNA and other RNA targets within organized biological specimens. Researchers can use those patterns to examine gene expression during development, distinguish cellular identities, study tissue organization, and investigate changes associated with disease. The observed location links RNA presence to specific biological structures and processes.