Fluorescent Probe Hybridization
Fluorescent Probe Hybridization - Related Videos
10:01
Observation and Quantification of Telomere and Repetitive Sequences Using Fluorescence In Situ Hybridization (FISH) with PNA Probes in Caenorhabditis elegans
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Cited by 5 •
2016
We report a concise procedure of fluorescence in situ hybridization (FISH) in the gonad and embryos of Caenorhabditis elegans for observing and quantifying repetitive sequences. We successfully observed and quantified two different repetitive sequences, telomere repeats and template of alternative lengthening of telomeres (TALT).
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.
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.
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.
12:31
Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization
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Cited by 136 •
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.
FAQs
Specificity comes from complementary base pairing between the labeled probe and its intended DNA or RNA sequence. A matching target supports probe annealing, whereas unrelated sequences are less likely to retain the probe under appropriate binding and washing conditions. This sequence dependence allows investigators to associate fluorescence with a particular nucleic-acid target rather than with cellular material generally.
Binding and washing conditions control the balance between detectable target recognition and retained probe interactions. Conditions must allow the probe to anneal to its complementary sequence while removing probe that has not remained associated with the target. Because fluorescence depends on what remains after washing, these controls directly affect whether a signal can be interpreted confidently.
The fluorescence pattern can show where the target occurs within a cell, tissue, or microbial sample. Sequence specificity links that location to a chosen DNA or RNA sequence. This combination helps investigators distinguish localized infection or expression patterns from a signal averaged across the entire specimen, adding spatial context to molecular detection.
A basic workflow begins by choosing a fluorescently labeled probe for the sequence of interest and applying it to the sample. Investigators then control probe-binding and washing conditions before examining the sample with a fluorescence microscope. The final image is interpreted by asking whether fluorescence appears and where it is located.
In infection studies, the signal can be used to localize pathogen nucleic acids within relevant samples and to distinguish infected cells from uninfected cells. That readout connects a molecular target with a cellular or tissue location, helping researchers characterize where infection is present rather than treating the sample as uniformly infected.
Within immunology, the technique can map immune-related gene expression to particular cells or regions while also revealing cellular responses associated with infection. This spatial context supports studies of host-pathogen interactions, allowing researchers to examine pathogen nucleic acids, immune-related expression, and cellular responses together within the sampled material.
Fluorescent Probe Hybridization can support diagnostic development by providing sequence-specific evidence in a visual format. It can also contribute to pathogen characterization when investigators need to determine whether a particular nucleic-acid target is present and where it appears in the sample. These uses connect molecular detection with infectious-disease research and assay development.