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首页> 外文会议>Smart Medical and Biomedical Sensor Technology II; Progress in Biomedical Optics and Imaging; vol.5, no.32 >Nanosphere-based SERS immune-sensors for protein analysis
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Nanosphere-based SERS immune-sensors for protein analysis

机译：基于纳米球的SERS免疫传感器用于蛋白质分析

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We have developed and optimized novel nanosphere-based silver coated SERS substrates for the detection of proteins. These SERS substrates were optimized for silver thickness, number of silver layers, and extent of silver oxidation between layers. Immuno-nanosensors capable of being inserted into individual cells and non-invasively positioned to the sub-cellular location of interest using optical tweezers were constructed from monodisperse silica nanospheres. Silica nanospheres ranging in diameter from 100 to 4500 nm were condensed from tetraalkoxysilanes in an alcoholic solution of water and ammonia. By varying the reaction conditions, accurate control of the silica nanospheres' diameter was achieved. Silica sphere sizes were optimized for SERS signal response. Nanosphere-based SERS substrates were made by depositing multiple layers of silver on the nanospheres, followed by binding of the antibody of interest to the silver. In binding the antibodies, different crosslinkers were characterized and compared. On one end, each of these crosslinkers contained sulfur or isothiocyanate groups which bound to the silver surface, while the other end contained a carboxylic or primary amine group which reacted readily with the antibodies. In order to evaluate these substrates, SERS spectra of different proteins, such as insulin and interleukin-2 (IL-2), were obtained. By using silver, as the metal surface for SERS, red and near-infrared excitation wavelengths (i.e., 600-700 nm) can be used. Excitation in this range helps to avoid photodamage to cells and reduces any autofluorescence background. Evaluation of these SERS substrates was performed using a 10 mW HeNe laser, operating at 632.8 nm, in a collinear excitation/detection geometry. The SERS signals were filtered with a holographic notch filter, dispersed by 1/3 meter spectrometer and detected using an intensified charge coupled device (ICCD). This paper discusses the fabrication and optimization of these nanosensors, as well as their potential applications.
机译：我们已经开发和优化了新型的基于纳米球的涂银SERS底物，用于检测蛋白质。这些SERS基板针对银的厚度，银的层数以及层之间的银氧化程度进行了优化。由单分散二氧化硅纳米球构建的免疫纳米传感器能够插入单个细胞并使用光镊无创地定位到感兴趣的亚细胞位置。在水和氨的酒精溶液中，将直径为100至4500 nm的二氧化硅纳米球与四烷氧基硅烷缩合。通过改变反应条件，可以精确控制二氧化硅纳米球的直径。硅胶球尺寸针对SERS信号响应进行了优化。通过在纳米球上沉积多层银，然后将目标抗体与银结合，来制备基于纳米球的SERS底物。在结合抗体时，表征和比较了不同的交联剂。一方面，这些交联剂中的每一个均包含键合至银表面的硫或异硫氰酸酯基，而另一端则包含易于与抗体反应的羧基或伯胺基。为了评估这些底物，获得了不同蛋白质（例如胰岛素和白介素2（IL-2））的SERS光谱。通过使用银作为SERS的金属表面，可以使用红色和近红外激发波长（即600-700nm）。在此范围内的激发有助于避免对细胞的光损伤并减少任何自发荧光背景。使用共线激发/检测几何形状的10 mW HeNe激光器（工作于632.8 nm）对这些SERS基板进行评估。 SERS信号用全息陷波滤波器过滤，用1/3米光谱仪分散，并使用增强电荷耦合器件（ICCD）进行检测。本文讨论了这些纳米传感器的制造和优化及其潜在应用。

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