Immunofluorescence assays
AssigneeVector Laboratories, Inc.
InventorsTimothy B. Karpishin, Pamela James, Erika Leonard
Life of this patent
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Abstract
Provided herein are immunofluorescence assays that do not use an intensely colored lipophilic dye, and that result in a significant reduction of fluorescence due to aldehyde fixation, collagen, elastin, and/or red blood cells. In certain embodiments of the present invention, a two-step treatment of the tissue is contemplated that leads to the incorporation of reduced heteropoly acids or salts thereof (e.g., phosphomolybdate species) in the tissue. This reagent binds tightly to hydrophilic regions of the tissue and effectively quenches the fluorescence in those regions. In doing so, the overall signal-to-noise ratio for the immunofluorescence assay is increased.
Claims14
A method for lowering/reducing/masking/suppressing tissue autofluorescence in an immunofluorescence assay, said method comprising, before or during conducting said immunofluorescence assay:
- providing a tissue comprising lung, liver, heart, nervous system, skin, lymph gland, musculoskeletal, spleen, eye, sinus, nasal mucosa, larynx, gastrointestinal tract tissue, reproductive organ, breast, prostate, salivary gland, tonsil, or kidney tissue;
- applying a heteropoly acid or salt thereof to said tissue, wherein the heteropoly acid comprises phosphomolybdic acid, phosphotungstic acid, silicomolybdic acid, silicotungstic acid, or a mixture of any two or more thereof or salts thereof;
- reducing at least a portion of said heteropoly acid or salt thereof to a reduced form with a reducing agent;
- and
- measuring the immunofluorescence of the tissue.
Dependencies are inferred from the claim text.