Effects of fluorophore-to-silver distance on the emission of cyanine-dye-labeled oligonucleotides

Joanna Malicka, Ignacy Gryczynski, Zygmunt Gryczynski, Joseph R. Lakowicz

Research output: Contribution to journalArticle

174 Citations (Scopus)

Abstract

We examined the fluorescence spectral properties of Cy3- and Cy5-labeled oligonucleotides at various distances from the surface of silver island films. The distance to the surface was controlled by alternating layers of biotinylated bovine serum albumin (BSA) and avidin, followed by binding of a biotinylated oligonucleotide. The maximum enhancement of fluorescence near a factor of 12 was observed for the first BSA-avidin layer, with the enhancement decreasing to 2-fold for six layers. The minimum lifetimes were observed for the first BSA-avidin layer, and were about 25-fold shorter than on quartz slides without silver, with the lifetimes being about 2-fold shorter for six BSA-avidin layers. These results suggest that maximum fluorescence enhancements occur about 90Å from the silver surface, a distance readily obtained by one or two layers of proteins.

Original languageEnglish
Pages (from-to)57-66
Number of pages10
JournalAnalytical Biochemistry
Volume315
Issue number1
DOIs
StatePublished - 1 Apr 2003

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Avidin
Fluorophores
Bovine Serum Albumin
Silver
Oligonucleotides
Coloring Agents
Fluorescence
Quartz
Islands
Proteins

Cite this

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abstract = "We examined the fluorescence spectral properties of Cy3- and Cy5-labeled oligonucleotides at various distances from the surface of silver island films. The distance to the surface was controlled by alternating layers of biotinylated bovine serum albumin (BSA) and avidin, followed by binding of a biotinylated oligonucleotide. The maximum enhancement of fluorescence near a factor of 12 was observed for the first BSA-avidin layer, with the enhancement decreasing to 2-fold for six layers. The minimum lifetimes were observed for the first BSA-avidin layer, and were about 25-fold shorter than on quartz slides without silver, with the lifetimes being about 2-fold shorter for six BSA-avidin layers. These results suggest that maximum fluorescence enhancements occur about 90{\AA} from the silver surface, a distance readily obtained by one or two layers of proteins.",
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Effects of fluorophore-to-silver distance on the emission of cyanine-dye-labeled oligonucleotides. / Malicka, Joanna; Gryczynski, Ignacy; Gryczynski, Zygmunt; Lakowicz, Joseph R.

In: Analytical Biochemistry, Vol. 315, No. 1, 01.04.2003, p. 57-66.

Research output: Contribution to journalArticle

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T1 - Effects of fluorophore-to-silver distance on the emission of cyanine-dye-labeled oligonucleotides

AU - Malicka, Joanna

AU - Gryczynski, Ignacy

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AU - Lakowicz, Joseph R.

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