Fluorescence Immunoassay Based on Long Time Correlations of Number Fluctuations

We report the development of a fluorescence-based immunoassay technique relying on the physical phenomena of random number fluctuations and diffusion, which we review. By determining the autocorrelation of the fluctuations in the fluorescent intensity, this method is able to measure the amount of la...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1980-08, Vol.77 (8), p.4904-4908
Main Authors: Nicoli, D. F., Briggs, J., Elings, V. B.
Format: Article
Language:English
Subjects:
Antibodies
Antigen-Antibody Complex
Autocorrelation
Charge carriers
Fluorescence
Fluorescent Antibody Technique
Immunoassay
Immunoassay - methods
Immunoglobulin G - analysis
Immunology
Molecules
Particle diffusion
Particle intensity
Particle motion
Spectrometry, Fluorescence
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Summary:We report the development of a fluorescence-based immunoassay technique relying on the physical phenomena of random number fluctuations and diffusion, which we review. By determining the autocorrelation of the fluctuations in the fluorescent intensity, this method is able to measure the amount of labeled antigen or antibody that is bound to micrometer-sized carrier particles in solution. The principal advantage of this technique is its insensitivity to small, fast-diffusing sources. It also discriminates against weakly fluorescent contaminants of size comparable to the carrier particles. We demonstrate these attributes by using two model systems: a human IgG assay and an idealized system consisting of polystyrene fluorescent spheres and rhodamine dye.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.77.8.4904