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. 2004 Jun 15;74(3):263-71.
doi: 10.1002/bip.20071.

Fluorescence spectral properties of labeled thiolated oligonucleotides bound to silver particles

Joanna Malicka  1 , Ignacy Gryczynski, Joseph R Lakowicz
Affiliations

Fluorescence spectral properties of labeled thiolated oligonucleotides bound to silver particles

Joanna Malicka et al. Biopolymers. .

Abstract

We examined the fluorescent spectral properties of fluorescein-labeled DNA oligomers when directly bound to metallic silver particles via a terminal sulfhydryl group. We found a 12-fold increase in fluorescence intensity and 25-fold decrease in lifetime for a fluorescein residue positioned 23 nucleotides from the silver surface compared to labeled oligomers in free solution. Similar results were found for a 23-mer labeled with five fluorescein residues. The absence of long lifetime components in the intensity decays suggests that all labeled oligomers are bound to silver and affected similarly by the metallic surfaces. These results provide the basic knowledge needed to begin use of metal-enhanced fluorescence for the detection of target sequences in simple formats potentially without a washing separation step. The use of metal-enhanced fluorescence provides a generic approach to obtaining a hybridization-dependent increase in fluorescence with most, if not all, commonly used fluorophores.

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Figures

FIGURE 1
FIGURE 1
Fluorescence emissions spectra of Fl-DNA-SH before (-) and after 48-h exposure (- - -) to a silver island film.
FIGURE 2
FIGURE 2
Fluorescence emission spectra of Fl-DNA-SH bound to a SIF (-) and compared to a roughly equivalent amount of Fl-DNA-SH in solution (- - -).
FIGURE 3
FIGURE 3
Frequency-domain intensity decays of Fl-DNA-SH in solution (top) and bound to a SIF (bottom).
FIGURE 4
FIGURE 4
Fluorescence emission spectra of Fl-DNA-SH on quartz and a SIF(s) when the amino groups were not blocked with succinic anhydride.
FIGURE 5
FIGURE 5
Frequency-domain intensity decays of Fl-DNA-SH on quartz (top), a SIF (middle) with unblocked amino groups, and a mixture of ds Fl-DNA-SH on SIFs and ss Fl-DNA in solution (bottom). In this mixture the steady-state intensities of the bound and unbound fluoresceinlabeled oligomers were roughly equal.
FIGURE 6
FIGURE 6
Emission spectra and frequency-domain intensity decays of Fl-DNA(Fl)4-SH in cuvette and on SIF with blocked amino groups.
FIGURE 7
FIGURE 7
Photostability of fluorescein-labeled oligomers bound to silver particles, the entire surface of silver island films (unblocked amino groups), or bound via a protein monolayer.
SCHEME 1
SCHEME 1
Chemical structure of the oligonucleotides.
SCHEME 2
SCHEME 2
Thiolated DNA bound to silver particles (top). Absorption spectrum of a SIF (left) and our experimental geometry (right).
SCHEME 3
SCHEME 3
Jablonski diagram for free fluorophores (top) and on SIFs (bottom).
See this image and copyright information in PMC

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