Enhanced fluorescence of Cy5-labeled DNA tethered to silver island films: fluorescence images and time-resolved studies using single-molecule spectroscopy.
Methods that increase the total emission per fluorophore would provide increased sensitivity and a wider dynamic range for chemical analysis, medical diagnostics, and in vivo molecular imaging. The use of fluorophore-metal interactions has the potential to dramatically increase the detectability of single fluorophores for bioanalytical monitoring. The fabrication and single-molecule analysis of fluorophore-labeled DNA molecules tethered to silver island films are described in this article. The single-molecule spectroscopic method reveals some insightful information on the behaviors of single molecules, rather than an ensemble of molecules. Analysis of fluorescence images, intensity profiles, total emitted photons, and lifetime distributions reveals some of sample heterogeneities. Investigations of time-dependent emission characteristics of single molecules indicate that the total number of emitted photons on the silvered surface is more than 10 times greater than on free labeled DNA molecules on a glass substrate. In addition, time-correlated single-photon counting results reveal the reduced lifetimes of single molecules tethered to silver island films.
Fu
Yi
Y
Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, MD 21201, USA.
Lakowicz
Joseph R
JR
D002232:Carbocyanines / Q000737:chemistry*; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D008856:Microscopy, Fluorescence / Q000379:methods*; D012834:Silver / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013997:Time Factors
D002232:Carbocyanines; C085321:cyanine dye 5; D012834:Silver; D004247:DNA