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Single Particle Raman Measurements of Gold Nanoparticles Used in Surface-Enhanced Raman Scattering (SERS)-Based Sandwich Immunoassays

机译:基于表面增强拉曼散射(SERS)的三明治免疫测定法中的金纳米颗粒的单颗粒拉曼测量

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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) from 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 ~70 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.
机译:已经研究了粒径对使用标记金纳米粒子的表面增强拉曼散射(SERS)强度的影响。进行了两组实验,均使用632.8 nm激光激发。第一种方法需要进行夹心免疫分析,其中将抗体与光滑的金底物偶联,从缓冲水溶液中选择性捕获游离前列腺特异性抗原(f-PSA)。然后,通过标称直径为30、40、50、60或80 nm的拉曼标记免疫金纳米颗粒的响应来检测捕获的f-PSA的存在。通过考虑颗粒表面积差异后计算每个颗粒的平均响应,将所得的SERS响应与颗粒密度相关联,这是通过原子力显微镜确定的。该分析表明,SERS响应的幅度随着粒径的增加而增加。第二组实验检查了单个纳米颗粒的响应。这些实验的不同之处在于,标记的纳米颗粒通过胺端基的硫醇盐偶联到光滑的金底物上,从而使颗粒和底物之间的平均间隔小得多。结果表明,直径约70 nm的颗粒表现出最大的增强作用。简要讨论了两组发现的差异的起因,这归因于纳米粒子和下面的基底之间的等离激元耦合的距离依赖性。

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