Optimizing Direct Immunofluorescence
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Immunofluorescence is a laboratory technique that utilizes a fluorophore-labeled antibody to detect immune complexes in tissue. Most of the labeled antibodies used in a clinical laboratory bind the conserved domains within each class of human antibodies, allowing them to detect a wide range of autoimmune complexes. Drawbacks to this technique mostly relate to proper handling of the specimen and the fluorophore-labeled antibodies. Therefore, having a basic understanding of fluorophores and antibodies is important for processing a specimen that yields a high signal-to-background ratio as well as troubleshooting problems, should they arise.
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