Fluorescence properties of labeled proteins near silver colloid surfaces

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Abstract
The fluorescence properties of a monolayer of labeled avidin molecules were studied near silver island films. We first adsorbed a monolayer of biotinylated‐BSA as a base that was used to capture labeled avidin molecules. For labeled avidin on silver island films, we observed an increase of the fluorescence intensity of between 18 and 80 with one‐photon excitation and up to several hundredfold or larger with two‐photon excitation. The probes were moderately more photostable in the presence of silver islands. There was also a dramatic decrease in the lifetimes with the amplitude‐weighted values decreasing from 7‐ to 35‐fold. The data suggest that these spectral changes are due to both increased rates of excitation near the metallic particles and increases in the rates of radiative decay. Because these silver island surfaces are very heterogeneous, we are hopeful that larger increases in intensity and photostability can be obtained for probes situated at an optimal distance from the ideal island surfaces. © 2003 Wiley Periodicals, Inc. Biopolymers (Biospectroscopy) 70: 585–594, 2003