Fluorescent In Situ Hybridization
Fluorescent In Situ Hybridization - Related Videos
12:31
Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization
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Cited by 133 •
2009
Whole mount in situ hybridization is one of the most widely used techniques in developmental biology. Here, we present a high-resolution double fluorescent in situ hybridization protocol for analyzing the precise expression pattern of a single gene and for determining the overlap of the expression domains of two genes. We include a propidium iodide nuclear counter-stain to highlight tissue organization.
16:19
Fluorescence in situ hybridization (FISH) Protocol in Human Sperm
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Cited by 39 •
2009
This video-article describes, step by step, how to process a semen sample to achieve good-quality fluorescence in situ hybridization on human spermatozoa. Preparations obtained can be used for aneuploidy screening in the context of clinical diagnosis.
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...
09:00
Fluorescent in situ Hybridization on Mitotic Chromosomes of Mosquitoes
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Cited by 45 •
2012
Among the three mosquito genera, namely Anopheles, Aedes, and Culex, physical genome mapping techniques were established only for Anopheles, whose members possess readable polytene chromosomes. For the genera of Aedes and Culex, however, cytogenetic mapping remains challenging because of the poor quality of polytene chromosomes. Here we present a universal protocol for obtaining high-quality preparations of mitotic chromosomes and an optimized FISH protocol for all three genera of mosquitoes.
03:42
Fluorescence In Situ Hybridization of Gut Bacteria in Mouse Intestinal Sections
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2026
Source: Ng, K. M., Tropini, C. Visualization of Gut Microbiota-host Interactions via Fluorescence In Situ Hybridization, Lectin Staining, and Imaging. J. Vis. Exp. (2021).This video demonstrates the procedure of fluorescence in situ hybridization (FISH) to stain gut microbiota within mouse intestinal tissue by hybridizing fluorescent probes to bacterial rRNA (ribosomal ribonucleic acid) and counterstaining the host structures for microscopic analysis.
FAQs
Denaturation converts target nucleic acids into single strands, exposing sequences that may be inaccessible in their paired form. A complementary fluorescently labeled probe can then bind through sequence matching. This order of events is essential because, without strand separation, the probe cannot efficiently recognize its target, and the resulting fluorescence would not reliably mark the intended location.
The probe sequence determines which DNA or RNA region can produce a fluorescent signal. Only a target containing the matching sequence should support probe binding, so probe selection connects the observed signal with a particular genetic sequence. This allows researchers to examine selected chromosome regions, gene-associated sequences, or RNA targets rather than viewing nucleic acids indiscriminately.
FISH can be directed toward DNA sequences or RNA sequences, depending on the target being examined. DNA-focused analyses support chromosome mapping, rearrangement detection, and copy-number assessment, whereas RNA-focused analyses can contribute to studying gene expression. In both cases, the signal remains interpretable within the surrounding cellular or tissue context.
FISH preserves the position of a detected sequence within a chromosome, cell, or tissue rather than reducing the result to sequence information alone. Fluorescence microscopy therefore links a genetic signal with physical organization and cellular context. That connection is valuable when researchers need to relate chromosome structure, gene location, expression, or microorganism distribution to biological surroundings.
A basic workflow begins by preparing the cells or tissue and exposing the target nucleic acids through denaturation. Fluorescently labeled probes are then allowed to hybridize with complementary sequences. After this sequence-recognition step, fluorescence microscopy is used to observe where signals occur and how they are distributed. The workflow preserves the relationship between molecular targets and their original cellular setting.
Researchers examine the location and distribution of fluorescent signals to evaluate selected genetic features. Signal positions can support chromosome mapping, while altered patterns may indicate gene rearrangements or copy-number changes. Because the signals are viewed within cells or tissues, interpretation can connect these sequence-level findings with chromosome structure and cellular organization rather than treating them as isolated molecular measurements.
FISH is useful when a study requires visual detection of selected DNA or RNA sequences in their biological setting. Applications include chromosome mapping, investigating gene rearrangements and copy-number changes, identifying microorganisms, and examining gene expression or cellular organization. Its broad value comes from combining sequence-specific detection with direct observation of where targets occur in cells or tissues.
For microorganism studies, probes can be selected to recognize nucleic acid sequences associated with the organisms of interest. Fluorescence microscopy then shows whether matching signals are present and where they occur within the examined material. This approach adds spatial information to detection, helping relate microorganism identity or distribution to the surrounding cells or tissue context.