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To perform immunofluorescence staining, a fluorophore must be conjugated (“tagged”) to an antibody. Staining procedures can be applied to both retained intracellular expressed antibodies, or to cell surface antigens on living cells. There are two general classes of immunofluorescence techniques: primary (direct) and secondary (indirect).
In the life sciences fluorescence microscopy is a powerful tool which allows the specific and sensitive staining of a specimen in order to detect the distribution of proteins or other molecules of interest. As a result, there is a diverse range of techniques for fluorescent staining of biological samples. [citation needed]
Staining and fluorescent tagging can serve similar purposes. Biological staining is also used to mark cells in flow cytometry, and to flag proteins or nucleic acids in gel electrophoresis. Light microscopes are used for viewing stained samples at high magnification, typically using bright-field or epi-fluorescence illumination.
Strong fluorescence when bound to DNA led to the rapid adoption of DAPI for fluorescent staining of DNA for fluorescence microscopy. Its use for detecting DNA in plant, metazoa and bacteria cells and virus particles was demonstrated in the late 1970s, and quantitative staining of DNA inside cells was demonstrated in 1977. Use of DAPI as a DNA ...
Fluorescence is widely used in the life sciences as a powerful and minimally invasive method to track and analyze biological molecules in real-time Some proteins or small molecules in cells are naturally fluorescent, which is called intrinsic fluorescence or autofluorescence (such as NADH, tryptophan or endogenous chlorophyll, phycoerythrin or ...
DyLight Fluor, a product line of fluorescent dyes; Fluorescein diacetate hydrolysis, a biochemistry laboratory test; Other dyes: Rhodamine, family of derivatives of xanthene used as dyes, indicators and fluorescent tracers; Methylene blue, blue thiazine dye also used as a medication; Haematoxylin, natural stain derived from hearthwood and used ...
Immunohistochemistry or IHC staining of tissue sections (or immunocytochemistry, which is the staining of cells), is perhaps the most commonly applied immunostaining technique. [2] While the first cases of IHC staining used fluorescent dyes (see immunofluorescence ), other non-fluorescent methods using enzymes such as peroxidase (see ...
Since then, Fluorescein was created as a fluorescent dye by Adolph von Baeyer in 1871 and the method of staining was developed and utilized with the development of fluorescence microscopy in 1911. [4] Ethidium bromide and variants were developed in the 1950s, [4] and in 1994, fluorescent proteins or FPs were introduced. [5]