Immunofluorescence
Immunofluorescence (pronounced: im•mu•no•fluor•es•cence) is a technique used for light microscopy with a fluorescence microscope and is used primarily on microbiological samples. This technique uses the specificity of antibodies to their antigen to target fluorescent dyes to specific biomolecule targets within a cell, and therefore allows visualization of the distribution of the target molecule through the sample.
Etymology
The term "Immunofluorescence" is derived from the words "Immuno", referring to the immune system, and "fluorescence", referring to the emission of light by a substance that has absorbed light or other electromagnetic radiation.
Technique
Immunofluorescence can be used in combination with other, non-antibody methods of fluorescent staining, for example, use of DAPI to label DNA. Several microscope designs can be used for analysis of immunofluorescence samples; the simplest is the conventional wide-field fluorescent microscope.
Types
There are two types of immunofluorescence techniques, direct and indirect.
- Direct Immunofluorescence - The direct immunofluorescence uses a single, primary antibody, chemically linked to a fluorophore. The primary antibody recognizes the target molecule and binds to a specific region called the epitope.
- Indirect Immunofluorescence - The indirect method uses two antibodies; the unlabeled first (primary) antibody specifically binds the target molecule, and the secondary, fluorophore-conjugated antibody, which recognizes the primary antibody and binds to it.
Related Terms
See Also
External links
- Medical encyclopedia article on Immunofluorescence
- Wikipedia's article - Immunofluorescence
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