Single-particle Raman measurements of gold nanoparticles used in surface-enhanced Raman scattering (SERS)-based sandwich immunoassays

The effect of particle size on the intensity of surface-enhanced Raman scattering (SERS) using labeled gold nanoparticles has been investigated. Two sets of experiments were preformed, both of which employed 632.8-nm laser excitation. The first entailed a sandwich immunoassay in which an antibody coupled to a smooth gold substrate selectively captured free-prostate specific antigen (f-PSA) front buffered aqueous solutions. The presence of captured f-PSA was then detected by the response of Raman-labeled immunogold nanoparticles with nominal diameters of 30, 40, 50, 60, or 80 nm. The resulting SERS responses were correlated to particle densities, which were determined by atomic force microscopy, by calculating the average response per particle after accounting for differences in particle surface area. This analysis showed that the magnitude of the SERS response increased with increasing particle size. The second set of experiments examined the response of individual nanoparticles. These experiments differed in that the labeled nanoparticles were coupled to the smooth gold substrate by an amine-terminated thiolate, yielding a much smaller average separation between the particles and substrate. The results revealed that particles with a diameter of similar to70 nm exhibited the largest enhancement. The origin of the difference in the two sets of findings, which is attributed to the distance dependence of the plasmon coupling between the nanoparticles and underlying substrate, is briefly discussed.