Metallic surfaces can have unusual effects on fluorophores such as increasing or decreasing the rates of radiative decay and the rates of resonance energy transfer (RET). In the present article we describe the effects of metallic silver island films on the emission spectra, lifetimes, and energy transfer for several fluorophores. The fluorophores are not covalently coupled to the silver islands so that there are a range of fluorophore-to-metal distances. We show that proximity of fluorophores to the silver islands results in increased fluorescence intensity, with the largest enhancement for the lowest-quantum-yield fluorophores. Importantly, the metal-induced increases in intensity are accompanied by decreased lifetimes and increased photostability. These effects demonstrate that the silver islands have increased the radiative decay rates of the fluorophore. For solvent-sensitive fluorophores the emission spectra shifted to shorted wavelengths in the presence of the silver islands, which is consistent with a decrease of the apparent lifetime for fluorophores near the metal islands. We also observed an increased intensity and blue spectral shift for the protein human glyoxalase, which displays a low quantum yield for its intrinsic tryptophan emission. In this case the blue shift is thought to be due to increased emission from a buried low-quantum-yield tryptophan residue. Increased intensities were also observed for the intrinsic emission of the nucleic acid bases adenine and thymine and for single-stranded 15-mers poly(T) and poly(C). And finally, we observed increased RET for donors and acceptors in solution and when bound to double-helical DNA. These results demonstrate that metallic particles can be used to modify the emission from intrinsic and extrinsic fluorophores in biochemical systems.

We examined the effects of metallic silver colloids on the fluorescence spectral properties of indocyanine green (ICG), which is a dye widely used for in vivo medical testing. Silver colloids from a suspension bind spontaneously to amine-coated surfaces. These colloid-coated surfaces were found to cause a 30-fold increase in the intensity of ICG, which was held close to the metal surface by adsorbed albumin. The increased intensities of ICG were also associated with decreased lifetimes and increased photostability, which are indicative of modifying the fluorophores radiative decay rate. These results suggest the use of metal colloid-enhanced ICG for applications to retinal angiography and vascular imaging and as a contrast agent for optical tomography.

We have used X-ray reflectivity, ellipsometry, and X-ray photoelectron spectroscopy to characterize self-assembled films of (3-aminopropyl)triethoxysilane (APS) functionalized with a photosensitive chromophore. Using the evolution of film thickness with time devoted to initial APS adsorption on the surface, we obtain a model for the structure and growth dynamics of the film. LA990557W(43) Dulcey, C.; Gru ¨neberg, K. Unpublished results.

We describe two reagentless methods of silver deposition for metal-enhanced fluorescence. Silver was deposited on glass positioned between two silver electrodes with a constant current in pure water. Illumination of the glass between the electrodes resulted in localized silver deposition. Alternatively, silver was deposited on an Indium Tin Oxide cathode, with a silver electrode as the anode. Both types of deposited silver produced a 5-18-fold increase in the fluorescence intensity of a nearby fluorophore, indocyanine green (ICG). Additionally, the photostability of ICG was dramatically increased by proximity to the deposited silver. These results suggest the use of silver deposited from pure water for surface-enhanced fluorescence, with potential applications in surface assays and lab-on-a-chip-based technologies, which ideally require highly fluorescent photostable systems.

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