Effects of metallic silver island films on resonance energy transfer between N,N'-(dipropyl)-tetramethyl- indocarbocyanine (Cy3)- and N,N'-(dipropyl)-tetramethyl- indodicarbocyanine (Cy5)-labeled DNA.
Resonance energy transfer (RET) is typically limited to distances below 60 A, which can be too short for some biomedical assays. We examined a new method for increasing the RET distances by placing donor- and acceptor-labeled DNA oligomers between two slides coated with metallic silver particles. A N,N'-(dipropyl)-tetramethylindocarbocyanine donor and a N,N'-(dipropyl)-tetramethylindodicarbocyanine acceptor were covalently bound to opposite 5' ends of complementary 23 base pair DNA oligomers. The transfer efficiency was 25% in the absence of silver particles or if only one slide was silvered, and it increased to an average value near 64% between two silvered slides. The average value of the Forster distance increased from 58 to 77 A. The energy transfer data were analyzed with a model assuming two populations of donor-acceptor pairs: unaffected and affected by silver island films. In an affected fraction of about 28%, the apparent energy transfer efficiency is near 87% and the Forster distance increases to 119 A. These results suggest the use of metallic silver particles to increase the distances over which RET occurs in biomedical and biotechnology assays.
Malicka
Joanna
J
University of Maryland Baltimore, Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201, USA. cfs@cfs.umbi.umd.edu
Gryczynski
Ignacy
I
Kusba
Jozef
J
Lakowicz
Joseph R
JR
D002232:Carbocyanines / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D011791:Quartz; D012834:Silver; D013194:Staining and Labeling
D002232:Carbocyanines; C089187:cyanine dye 3; C085321:cyanine dye 5; D011791:Quartz; D012834:Silver; D004247:DNA