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Immunofluorescence is an assay which is used primarily on biological samples and is
classically defined as a procedure to detect antigens in cellular contexts using antibodies.
The specificity of antibodies to their antigen is the base for immunofluorescence.
The biological samples include tissue and cells.
Immunofluorescence allows researchers to evaluate whether or not cells in a particular sample
express the antigen in question.
In cases where an immune positive signal is found, immunofluorescence also allows
researchers to determine which subcellular compartments are expressing the antigen.
Immunofluorescence can be used on cultured cell lines, tissue sections, or individual cells.
Immunofluorescence may be used to analyse the distribution of proteins, glycans, and small
biological and non-biological molecules.
Immunofluorescence has been widely used in biological research and medical research yield
and becomes one most important and effective method.
TWO DIFFERENT IMMUNOFLUORESCENCE ASSAY
The antibody is itself conjugated with the fluorochrome and applied directly to
a monolayer of cells or to frozen tissue on a slide.
When examined with a fluorescence microscope, the antibody labeled with the
fluorescent compound identifies the localized antigen.
Unlike direct immunofluorescence, indirect immunofluoresence is a double-layer technique.
The unlabeled antibody is applied directly to the tissue substrate and then treated with a
There are several advantages to this technique, and it is typically used more frequently than the
Because several fluorescent anti-immunoglobulins can bind to each antibody present in the
first layer, this produces brighter fluorescence than in the direct method.
It is also more time-efficient since there is only one fluorescent-labeled reagent, the anti-IgG
prepared during the lengthy conjugation process.