Immunofluorescence detection methods using microspheres

Ferenc Szurdoki; Karri L. Michael; Divya Agrawal; Laura C. Taylor; Sandra L. Schultz; David R. Walt

doi:10.1117/12.335781

12 January 1999 Immunofluorescence detection methods using microspheres

Ferenc Szurdoki; Karri L. Michael; Divya Agrawal; Laura C. Taylor; Sandra L. Schultz; David R. Walt

Author Affiliations +

Proceedings Volume 3544, Pathogen Detection and Remediation for Safe Eating; (1999) https://doi.org/10.1117/12.335781
Event: Photonics East (ISAM, VVDC, IEMB), 1998, Boston, MA, United States

Abstract

Microsphere-based immunoassays were devised for compounds of agricultural and biomedical interest (e.g., digoxin, theophylline, and zearalenone). Commercially available microspheres with surface functional groups for chemical derivatization were used as solid carriers. After immobilizing the target substances, the surface of the haptenized microspheres was blocked by a protein to reduce aspecific binding. Competitive immunoassays were performed using the functionalized microspheres and antibodies labeled with horseradish peroxidase. Immunofluorescence signal amplification was achieved by enzyme-catalyzed reporter deposition (CARD). An epifluorescence microscope, a CCD camera interfaced with a computer, and microscopy image analysis software were employed for quantitative detection of fluorescent light emitted from individual microspheres. Integration of several such immunoassays and application of an optical encoding method enabled multianalyte determination. These immunoassays can also be utilized in an immunosensor array format. This immunoarray format could facilitate miniaturization and automation of multianalyte immunoassays.

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Ferenc Szurdoki, Ferenc Szurdoki, Karri L. Michael, Karri L. Michael, Divya Agrawal, Divya Agrawal, Laura C. Taylor, Laura C. Taylor, Sandra L. Schultz, Sandra L. Schultz, David R. Walt, David R. Walt, } "Immunofluorescence detection methods using microspheres", Proc. SPIE 3544, Pathogen Detection and Remediation for Safe Eating, (12 January 1999); doi: 10.1117/12.335781; https://doi.org/10.1117/12.335781

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