Immobilization of nanoparticles (NPs) is a technique suitable for the preparation of large-scale substrates for surface-enhanced vibrational spectroscopy including micro- and nano-spectroscopic applications. The developed immobilization method provides the enhancing properties of the roughened substrate surface to be maintained for techniques like surface-enhanced Raman scattering (SERS) spectroscopy, however, at the same time the morphology is not limiting for related near-field (scanning probe) techniques. The study is focused on the comparison of different immobilization procedures of Ag nanoparticles and finding the relationship between preparation procedures leading to convenient surface morphology and the quality of the observed signal of the model analyte (riboflavin) using SERS. Amino-linker (3-aminopropyl)trimethoxysilane (APTMS) and four thio-linkers (cysteine, 3-mercaptopropanoic acid, 2-mercaptoethanol, and 2,2′-oxydiethanthiol) using five immobilization procedures at three different temperatures (23 °C, 40 °C, and 70 °C) were compared. Surface morphology was monitored by scanning electron microscopy and atomic force microscopy. The SERS spectra of riboflavin were evaluated in terms of the intensity and the resolution of individual bands. The spectral dataset was inspected by multivariate statistical methods - principal component analysis and discriminant analysis. The evaluation of spectra and statistical models show the influence of the used linker and AgNPs immobilization procedure on the spectral output. APTMS linker is less suitable; much more appropriate are thio-linkers deposited on an evaporated Au layer on a glass slide. The best spectral parameters were obtained for 2,2′-oxydiethanthiol and 23 °C.

纳米颗粒的固定化（NPs）是一种适用于制备大型基质的技术，用于表面增强振动光谱学，包括微光谱和纳米光谱应用。研发的固定方法为表面增强拉曼散射（SERS）光谱等技术提供了要保持的粗糙基材表面的增强性能，但是，同时形态并不局限于相关的近场（扫描探针）技术。这项研究的重点是比较不同的银纳米颗粒固定方法的比较，并发现可方便表面形态的制备程序与使用SERS观察到的模型分析物（核黄素）信号质量之间的关系。在三个不同的温度（23°C，23°C）下采用五种固定步骤，使用了五个连接步骤的氨基连接基（3-氨基丙基）三甲氧基硅烷（APTMS）和四个硫基连接基（半胱氨酸，3-巯基丙酸，2-巯基乙醇和2,2'-氧二硫醇）。比较40°C和70°C）。通过扫描电子显微镜和原子力显微镜来监测表面形态。核黄素的SERS谱根据强度和各个谱带的分辨率进行了评估。通过多元统计方法-主成分分析和判别分析检查光谱数据集。光谱和统计模型的评估显示了所用的接头和AgNPs固定程序对光谱输出的影响。APTMS链接器不太适合；更合适的是在玻璃载片上的蒸镀金层上沉积硫键。对于2,2'-氧二硫醇和23°C，可获得最佳光谱参数。