Multi-analyte multi-color enhanced fluorescent immunoassay on silver island-coated surface

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

Research output: Contribution to journalConference articlepeer-review

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.

Original languageEnglish
Article number65
Pages (from-to)366-374
Number of pages9
JournalProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume5692
DOIs
StatePublished - 21 Jul 2005
EventAdvanced Biomedical and Clinical Diagnostic Systems III - San Jose, CA, United States
Duration: 23 Jan 200526 Jan 2005

Keywords

  • Enhanced fluorescence
  • Fluorescence immunoassay
  • Immunoglobulin
  • Multi-analyte
  • Multi-color
  • Nanosize metallic particles
  • Silver island films

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