Depolarization of surface-enhanced fluorescence: an approach to fluorescence polarization assays.
Localized surface plasmons of metallic particles of subwavelength sizes strongly modify the spectral properties of nearby fluorophores. The enhanced radiative decay rate leads to high fluorescence efficiencies and decreased fluorescence lifetimes. In this report we show that metal-enhanced fluorescence generated by the presence of the silver islands on the glass substrate displays high depolarization. Intensities, lifetimes, and emission anisotropies of several fluorophore protein conjugates have been studied in the absence and presence of metallic nanostructures. Despite highly decreased lifetimes of about 10-fold and immobilization of conjugates on the solid substrate, the observed emission anisotropies for all fluorophores on the metal-enhanced substrate decreased 300-500% compared to that in solution. This observation implies a new generation of fluorescence polarization immunoassays with broad applications because of no restrictions to the lifetime of the probe and the size of labeled biomolecules. The changes in polarization are due to binding that occur on the bioactive surface localized near the metal particles.
Szmacinski
Henryk
H
Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, University of Maryland Baltimore, 725 West Lombard Street, Baltimore, Maryland 21201, USA. henry@cfs.umbi.umd.edu
Lakowicz
Joseph R
JR
D004735:Energy Transfer; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes / Q000737:chemistry*; D012834:Silver / Q000737:chemistry*; D019809:Streptavidin / Q000737:chemistry*; D020349:Surface Plasmon Resonance
D005456:Fluorescent Dyes; D012834:Silver; D019809:Streptavidin