Fluorescence Hybridization
Fluorescence Hybridization - Related Videos
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.
12:15
Double Fluorescence in situ Hybridization in Fresh Brain Sections
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Cited by 7 •
2010
This protocol involves a non-radioactive in-situ hybridization procedure that enables the simultaneous identification of two transcript species, at a single cell resolution, in thin sections of the vertebrate brain.
FAQs
Denaturation separates the paired strands of target DNA or RNA, making their sequences accessible to the fluorescently labeled probes. During the subsequent annealing step, complementary sequences pair. This sequence-specific pairing is central to accurate localization, because fluorescence appears where the probe has bound its matching target within the cell or tissue.
The probe’s complementary sequence determines which DNA or RNA target can be recognized, while its fluorescent label supplies the visible readout. After hybridization, fluorescence microscopy shows the probe’s cellular or tissue position. This positional information links a selected molecular sequence with its location in the biological sample.
The observed signal pattern can indicate how a target sequence is represented or arranged in a sample. In cancer research, these patterns are examined for gene amplification, deletions, translocations, and broader chromosomal abnormalities. Interpreting the pattern at the cellular level helps connect a genomic change with the tumor cells in which it occurs.
A typical workflow begins with a cell or tissue sample, followed by denaturation of the target nucleic acids and probes. The complementary strands are then allowed to anneal, and the sample is examined by fluorescence microscopy. The resulting probe locations and signal patterns provide the basis for identifying sequence-specific genomic changes.
The essential components are a cell or tissue sample, nucleic acid probes carrying fluorescent labels, conditions that permit denaturation and complementary annealing, and a fluorescence microscope. Each serves a distinct role: the sample supplies the genetic material, the probe marks the selected sequence, and microscopy makes the resulting localization visible for analysis.
In cancer research, investigators apply it to tumor samples when they need to examine gene amplification, deletions, translocations, or chromosomal abnormalities at the cellular level. The findings can contribute to cancer classification and biomarker analysis, and they may support diagnosis and treatment planning by revealing genomic changes directly within the tumor material.