Increased resonance energy transfer between fluorophores bound to DNA in proximity to metallic silver particles.
We examined the effects of metallic silver particles on resonance energy transfer (RET) between fluorophores covalently bound to DNA. A coumarin donor and a Cy3 acceptor were positioned at opposite ends of a 23-bp double helical DNA oligomer. In the absence of silver particles the extent of RET is near 9%, consistent with a Forster distance R(0) near 50 A and a donor to acceptor distance near 75 A. The transfer efficiency increased when the solution of AMCA-DNA-Cy3 was placed between two quartz plates coated with silver island films to near 64%, as determined by both steady-state and time-resolved measurements. The apparent R(0) in the presence of silver island films increases to about 110 A. These values of the transfer efficiency and R(0) represent weighted averages for donor-acceptor pairs near and distant from the metallic surfaces, so that the values at an optimal distance are likely to be larger. The increased energy transfer is observed only between two sandwiched silvered slides. When we replaced one silvered slide with a quartz plate the effect vanished. Also, the increased energy transfer was not observed for silvered slides separated more than a few micrometers. These results suggest the use of metal-enhanced RET in PCR, hybridization, and other DNA assays, and the possibility of controlling energy transfer by the distance between silver surfaces.
Malicka
Joanna
J
University of Maryland Baltimore, Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, 725 W Lombard Street, Baltimore, MD 21201, USA.
Gryczynski
Ignacy
I
Fang
Jiyu
J
Kusba
Jozef
J
Lakowicz
Joseph R
JR
D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D004735:Energy Transfer; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes / Q000737:chemistry*; D012834:Silver / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; D012834:Silver; D004247:DNA