Rna Fluorescence In Situ Hybridization
Rna Fluorescence In Situ Hybridization - Related Videos
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.
04:23
Determining Viral RNA Distribution in Host Cells by Fluorescence In Situ Hybridization
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2026
Source: Vallery, T. K. & Steitz, J. A. Quantitative Fluorescence In Situ Hybridization (FISH) and Immunofluorescence (IF) of Specific Gene Products in KSHV-Infected Cells. J. Vis. Exp. (2019)This video demonstrates fluorescence in situ hybridization to visualize viral RNA in virus-infected cells. It outlines the steps involved in probe hybridization, signal amplification, and confocal imaging.
02:59
Using Immuno-RNA Fluorescence In Situ Hybridization to Visualize SARS-CoV-2
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2025
This video demonstrates the application of Hybridization Chain Reaction (HCR)-Fluorescence In Situ Hybridization (FISH) for visualizing the spatial distribution of SARS-CoV-2 subgenomic RNA. Combining HCR with immunofluorescence techniques has the potential to provide insights into host-virus interactions at the single-cell level.
13:06
Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization (FISH)
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Cited by 6 •
2012
Fluorescent in situ hybridization (FISH) to identify mRNA transcripts in individual cells allows analysis of polygenic activity such as the simultaneous transcription of more than one member of the var multigene family in Plasmodium falciparum infected erythrocytes 1. The technique is adaptable and can be used on different types of genes, cells and organisms.
FAQs
Probe sequence complementarity provides the recognition step: each fluorescently labeled probe binds its matching RNA sequence within the prepared sample. The resulting fluorescence marks the position of transcripts rather than merely indicating that a gene is active somewhere in the specimen. This sequence-directed signal supports spatial interpretation of developmental gene expression.
Fixation and permeabilization serve different purposes in the workflow. Fixation preserves the cells or tissue in place, maintaining the spatial relationships that matter for developmental analysis. Permeabilization then allows probes to access RNA inside the preserved sample. Together, these preparation steps make intracellular transcript localization compatible with fluorescence microscopy.
Multiplex RNA fluorescence in situ hybridization extends the analysis by detecting several transcripts in the same sample. Comparing their fluorescent patterns can show whether genes occupy shared or distinct regions, helping connect transcript distribution with cell identity, developmental signaling, and tissue organization. This is especially useful when several expression patterns must be interpreted together.
A basic RNA fluorescence in situ hybridization workflow starts with a fixed cell or tissue sample, followed by permeabilization, application of fluorescently labeled probes, and hybridization to complementary RNA sequences. The prepared material is then examined by fluorescence microscopy. This sequence links sample preservation and probe binding to the final spatial readout of transcript distribution.
In developmental biology, the method can compare expression patterns across stages of cell differentiation, tissue patterning, or embryonic development. Researchers can ask where a transcript appears and how that location changes as development proceeds. Such spatial comparisons help relate gene expression to the emergence of distinct cell identities and organized tissue structures.
The principal outcome is a spatial map of target RNA within the imaged cells or tissue. These maps retain information about gene-expression location that would be lost if analysis considered expression without its tissue context. When multiple transcripts are measured together, the combined patterns provide a basis for interpreting relationships among signaling, identity, and tissue organization.