Rna In Situ Hybridization
Rna 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...
09:57
Whole Mount RNA Fluorescent in situ Hybridization of Drosophila Embryos
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Cited by 22 •
2013
Here we describe a whole-mount fluorescent in situ hybridization (FISH) protocol for determining the expression and localization properties of RNAs expressed during embryogenesis in the fruit fly, Drosophila melanogaster.
10:16
Detection of Viral RNA by Fluorescence in situ Hybridization (FISH)
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Cited by 35 •
2012
A fluorescence in situ hybridization (FISH) method was developed to visually detect viral genomic RNA using fluorescence microscopy. A probe is made with specificity to the viral RNA that can then be identified using a combination of hybridization and immunofluorescence techniques. This technique offers the advantage of identifying the localization of the viral RNA or DNA at steady-state, providing information on the control of intracellular virus trafficking events.
03:26
Probe Hybridization and Signal Amplification in RNA In Situ Hybridization: A Technique for Detecting Specific RNA Sequences in Tissue Sections
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2023
This video describes a sequential hybridization strategy that involves hybridizing mRNA probes to specific target mRNA. Following probe hybridization, signal amplification is performed to enhance resolution via reduction of signal-to-noise ratio during RNA-CISH of histological samples.
08:13
RNA In situ Hybridization in Whole Mount Embryos and Cell Histology Adapted for Marine Elasmobranchs
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Cited by 1 •
2013
By combining methods for RNA whole mount in situ hybridization and histology, gene expression can be linked with cell fate decisions in the developing embryo. These methods have been adapted to marine elasmobranchs and facilitate the use of these animals as model organisms for biomedical, toxicology and comparative studies.
FAQs
Selectivity comes from complementary base pairing between a labeled RNA or DNA probe and its target transcript. Only sequences with sufficient complementarity retain the probe as the assay proceeds, allowing the resulting signal to indicate the location of that RNA rather than RNA broadly. This sequence-based recognition makes it possible to distinguish expression patterns associated with particular genes.
Both detection strategies reveal where a probe has bound, but they produce different signal types. Fluorescent labels generate light-based signals, whereas enzymatic labels produce signals through an enzyme-driven reaction. The choice affects how the detected transcript is visualized and can support different imaging approaches for examining expression within preserved cells or tissue.
Spatial information connects transcript presence with biological organization. Instead of indicating only that an RNA exists in a sample, the pattern shows which cell types or tissue regions contain it and how expression relates to tissue architecture. This context is especially valuable when expression changes across developmental stages or among distinct cellular environments.
The assay begins with preserved cells or tissue, followed by exposure to a labeled complementary RNA or DNA probe. Sequence-specific binding identifies locations containing the target transcript, after which the bound probe is visualized through a fluorescent or enzymatic signal. The resulting pattern is interpreted in relation to cell type, tissue structure, or developmental stage.
In developmental studies, the method can reveal how transcript locations change across embryonic stages and within developing tissues. Researchers can relate these patterns to emerging tissue organization and particular cell types, providing spatial context for gene expression during development. This makes it useful when the timing and location of expression are both biologically important.
Mapping transcripts within preserved tissue allows expression patterns to be compared with tissue architecture and cellular arrangement. In disease-focused work, the same spatial readout can show where disease-associated expression changes occur rather than treating the tissue as a uniform sample. These observations help connect molecular changes with specific regions or cell populations.
Multiplexed versions enable simultaneous analysis of multiple transcripts, allowing researchers to examine several expression patterns within the same spatial context. High-resolution formats provide more detailed localization within cells or tissue structures. Together, these extensions improve the ability to compare transcripts, identify relationships among expression domains, and study complex cellular responses.