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Immunoassays based on directional surface plasmon-coupled emission.

Evgenia Matveeva1, Zygmunt Gryczynski, Ignacy Gryczynski

  • 1Department of Biochemistry and Molecular Biology, Center for Fluorescence Spectroscopy, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, MD 21201, USA. eva@cfs.umbi.umd.edu

Journal of Immunological Methods
|April 17, 2004
PubMed
Summary

Surface plasmon-coupled emission (SPCE) enhances immunoassay sensitivity by directing fluorescence into a beam. This novel approach improves light collection and background suppression for more precise detection in assays.

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Area of Science:

  • Biophysics
  • Analytical Chemistry
  • Materials Science

Background:

  • Fluorescence-based assays are limited by isotropic light emission and inefficient collection.
  • Surface plasmon-coupled emission (SPCE) offers a method to enhance light collection efficiency.
  • SPCE involves coupling excited fluorophores with a thin metal film to direct emission.

Purpose of the Study:

  • To introduce and validate a novel immunoassay approach using SPCE.
  • To demonstrate the capability of SPCE for sensitive and selective detection.
  • To explore the potential of SPCE in surface-bound assays and microfluidic systems.

Main Methods:

  • Developed a model affinity assay using SPCE with labeled goat anti-rabbit IgG antibodies.
  • Utilized a 50-nm-thick silver film for surface plasmon coupling.

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  • Measured increased fluorescence intensity at a specific angle (75 degrees) upon antibody binding.
  • Investigated excitation under surface plasmon resonance (SPR) conditions.

    • Main Results:

    • SPCE efficiently collected and directed fluorescence emission into a cone-like beam.
    • Binding of labeled IgG to the silver film resulted in a significant increase in SPCE intensity.
    • SPCE intensity was dependent on fluorophore proximity to the silver film.
    • Background suppression was achieved as only nearby fluorophores contributed to SPCE.

    Conclusions:

    • SPCE provides a new technology for highly sensitive and selective surface-bound assays.
    • The method does not require changes in quantum yield upon binding.
    • SPCE offers advantages in light collection efficiency and background suppression compared to conventional methods.
    • Potential applications include microfluidic systems and advanced diagnostic tools.