Multi-analyte multicolor enhanced fluorescent immunoassay on silver island-coated surface

Evgenia G. Matveeva; Zygmunt Karol Gryczynski; Joseph R. Lakowicz

doi:10.1117/12.591685

1 April 2005 Multi-analyte multicolor enhanced fluorescent immunoassay on silver island-coated surface

Evgenia G. Matveeva, Zygmunt Karol Gryczynski, Joseph R. Lakowicz

Proceedings Volume 5692, Advanced Biomedical and Clinical Diagnostic Systems III; (2005) https://doi.org/10.1117/12.591685
Event: SPIE BiOS, 2005, San Jose, CA, United States

Abstract

A multi-analyte multi-color immunoassay utilizing enhanced fluorophores fluorescence in the proximity of silver island film (SIF) coated surface is described. A mixture of four different model analyte proteins was immobilized on the surface, and fluorescence signal of four corresponding antibodies labeled with different color fluorophores (emission maxima varied from 570 to 700 nm) was monitored using appropriate cut-off filters. To localize the excitation volume within close proximity of the SIF surface we used the total internal reflection (TIR) mode of excitation. Use of SIF coating resulted in approximately 5-10 fold increase in the intensity of the fluorophores signal if compared to the non-coated glass substrate surface. We suggest application of this methodology for multiplex immunoassays for simultaneous detection of several different analytes in medicine and biotechnology.

Citation Download Citation

Evgenia G. Matveeva, Zygmunt Karol Gryczynski, and Joseph R. Lakowicz "Multi-analyte multicolor enhanced fluorescent immunoassay on silver island-coated surface", Proc. SPIE 5692, Advanced Biomedical and Clinical Diagnostic Systems III, (1 April 2005); https://doi.org/10.1117/12.591685

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