In situ hybridization

In situ hybridization

In situ hybridization (ISH) is a type of hybridization that uses a labeled complementary DNA or RNA strand (i.e., probe) to localize a specific DNA or RNA sequence in a portion or section of tissue ("in situ"), or, if the tissue is small enough (e.g. plant seeds, Drosophila embryos), in the entire tissue (whole mount ISH). This is distinct from immunohistochemistry, which localizes proteins in tissue sections. DNA ISH can be used to determine the structure of chromosomes. Fluorescent DNA ISH (FISH) can, for example, be used in medical diagnostics to assess chromosomal integrity. RNA ISH (hybridization histochemistry) is used to measure and localize mRNAs and other transcripts within tissue sections or whole mounts.

Process

For hybridization histochemistry, sample cells and tissues are usually treated to fix the target transcripts in place and to increase access of the probe. As noted above, the probe is either a labeled complementary DNA or, now most commonly, a complementary RNA (riboprobe). The probe hybridizes to the target sequence at elevated temperature, and then the excess probe is washed away (after prior hydrolysis using RNase in the case of unhybridized, excess RNA probe). Solution parameters such as temperature, salt and/or detergent concentration can be manipulated to remove any non-identical interactions (i.e. only exact sequence matches will remain bound). Then, the probe that was labeled with either radio-, fluorescent- or antigen-labeled bases (e.g., digoxigenin) is localized and quantitated in the tissue using either autoradiography, fluorescence microscopy or immunohistochemistry, respectively. ISH can also use two or more probes, labeled with radioactivity or the other non-radioactive labels, to simultaneously detect two or more transcripts.

References

#Jin L, Lloyd RV. In situ hybridization: methods and applications. J Clin Lab Anal. 11(1):2-9, 1997. PMID 9021518
# [http://intramural.nimh.nih.gov/lcmr/snge/Protocols/ISHH/ISHH.html Comprehensive and annotated in situ hybridization histochemistry]

External links

*
* [http://www.cshprotocols.org/cgi/content/full/2008/3/pdb.prot4944 Whole-Mount In Situ Hybridization of RNA Probes to Plant Tissues]
* [http://www.cshprotocols.org/cgi/content/full/2007/10/pdb.prot4730 Preparation of Complex DNA Probe Sets for 3D FISH with up to Six Different Fluorochromes]
* [http://www.cshprotocols.org/cgi/content/full/2006/21/pdb.prot4519 Transcript In Situ Hybridization of Whole-Mount Embryos for Phenotype Analysis of RNAi-Treated Drosophila]


Wikimedia Foundation. 2010.

Игры ⚽ Нужно сделать НИР?

Look at other dictionaries:

  • In situ hybridization — The use of a DNA or RNA probe to detect the complementary sequence. In situ hybridization is like all nucleic acid hybridization in being a technique in which single stranded nucleic acids (DNA or RNA) are permitted to interact so that complexes …   Medical dictionary

  • in situ hybridization — hibridizacija in situ statusas T sritis augalininkystė apibrėžtis Specifinių DNR sekų atpažinimo metodas sveikose chromosomose su radioaktyviu papildomuoju (komplementariuoju) zondu. atitikmenys: angl. in situ hybridization rus. гибридизация in… …   Žemės ūkio augalų selekcijos ir sėklininkystės terminų žodynas

  • in situ hybridization —  In situ hybridization  Гибридизация in situ   Метод, использующий одноцепочечный РНК или ДНК зонд для обнаружения гена или молекулы информационной РНК в клетке или ткани (см. также гибридизация) …   Толковый англо-русский словарь по нанотехнологии. - М.

  • in-situ hybridization — noun (biology) A technique, using a radioactive nucleic acid probe and autoradiography, for locating the position of a specific DNA sequence on a chromosome • • • Main Entry: ↑in situ …   Useful english dictionary

  • in situ hybridization — in situ hybridization. См. цитологическая гибридизация. (Источник: «Англо русский толковый словарь генетических терминов». Арефьев В.А., Лисовенко Л.А., Москва: Изд во ВНИРО, 1995 г.) …   Молекулярная биология и генетика. Толковый словарь.

  • in situ hybridization — (1) Technique for revealing patterns of gene expression in a tissue. The tissue is fixed and prepared (usually by sectioning) on a slide, and a labelled DNA or RNA probe is hybridized to the sample. It binds only to complementary mRNA sequences,… …   Dictionary of molecular biology

  • Fluorescence in situ hybridization — A metaphase cell positive for the bcr/abl rearrangement (associated with chronic myelogenous leukemia) using FISH. The chromosomes can be seen in blue. The chromosome that is labeled with green and red spots (upper left) is the one where the… …   Wikipedia

  • Fluorescent in situ hybridization — can be used to find out where the fluorescent probe bound to the chromosome. FISH is often used for finding specific features in DNA. These features can be used in genetic counseling, medicine, and species identification. Probes Probes are often… …   Wikipedia

  • Fluorescent in situ hybridization — Hybridation fluorescente in situ Pour les articles homonymes, voir FISH. Technique de l hybridation fluorescente in situ. En A : sonde. B : sonde colorée à l aide d un …   Wikipédia en Français

  • Fluorescent in situ hybridization — An important molecular cytogenetic method for identifying chromosomes and parts of chromosomes, deciphering chromosome rearrangements, and locating genes on chromosomes. Fluorescent means emitting light that comes from a reaction within the… …   Medical dictionary

Share the article and excerpts

Direct link
… Do a right-click on the link above
and select “Copy Link”