Ultra-High-Throughput Screening (uHTS) Based on Surface*

描述（由申请人提供）：本研究的目的是开发基于表面增强拉曼散射（SERS）检测的新一代超高通量筛选（uHTS）系统。该项目的具体目标是：目标1。开发一种多光谱拉曼系统（MRS）作为超高温超导的光学读出器。目的2：评价利用分子哨兵进行高度平行配体/靶标结合检测的uHTS-MRS。目标3：开发和集成一个自动化系统，用于检测基于sers的纳米探针的光信号。目的4：评估CMOS-uHTS系统在细胞模型中高通量筛选小分子或mRNA探针的能力。我们的系统将为标记多路复用提供一个新的维度，用于在基于细胞的高通量筛选药物发现中表征大量样品。一种新的基于sers的分子前哨检测方案将应用于多重基因表达分析。该系统的独特之处包括：基于多光谱成像检测的新型仪器，基于分子识别和SERS检测相结合的新型检测方案，SERS效应将传统拉曼信号的灵敏度提高了106-1015倍，SERS- ms纳米探针对光漂白（通常与荧光探针相关）不敏感。超高通量“标签复用”检测（即同时使用许多拉曼染料标记探针的能力）（极窄的拉曼波段允许SERS用作复用技术）。该系统将用于筛选小分子、合成化学和天然产物文库，这些文库可以上调或下调各种mRNA家族的表达。能够调节mRNA基因表达水平的活性化合物将被鉴定，随后进行额外的分析。基于sers的uHTS仪器将提供一种比现有系统更快、更有效的潜在工具，并且能够提供高灵敏度、更高水平的特异性和多路复用能力，以筛选合成化学和天然产物库，例如那些将在美国国立卫生研究院资助的分子库筛选中心注册和容纳的化学和天然产物库。

DESCRIPTION (provided by applicant): The objective of this research is to develop a new generation of ultra-high-throughput screening (uHTS) systems based on surface-enhanced Raman scattering (SERS) detection. The specific aims of the project are: Aim 1. Develop a multi-spectral Raman system (MRS) as the optical readout for uHTS. Aim 2: Evaluate the uHTS-MRS using molecular sentinels for highly parallel ligand/target binding detection. Aim 3: Develop and integrate an automated system for detection of optical signals from SERS-based nanoprobes. Aim 4: Evaluate the CMOS-uHTS system for high throughput screening of small molecules or mRNA probes in a cellular model. Our system will offer a new dimension in label multiplexing for characterizing large numbers of samples in cell-based high-throughput screening for drug discovery. A novel SERS-based molecular-sentinel detection scheme will be applied for multiplex gene expression analysis. The unique features of the proposed system include: Novel instrumentation based on multi-spectral imaging detection Novel detection scheme based on combination of molecular recognition and SERS detection The SERS effect enhances the sensitivity of conventional Raman signal over 106-1015 fold SERS-MS nanoprobes are insensitive to photo-bleaching (often associated with fluorescent probes). Ultra high-throughput "label multiplex" detection (i.e. capability for many Raman dye-labeled probes to be used simultaneously) (the extremely narrow Raman bands allow SERS to be used as a multiplexing technique). The system will be tested for screening of small molecules, synthetic chemical and natural product libraries that up- or down-regulate expression of various families of mRNA. Active compounds that are capable of modulating mRNA gene expression levels will be identified, followed by additional assays. The SERS-based uHTS instrumentation will provide a tool that has the potential to be faster and more efficient than currently available systems, and is capable to provide great sensitivity with higher levels of specificity, and multiplexing capabilities to screen synthetic chemical and natural product libraries such as the ones that will be registered and housed in the NIH-sponsored molecular libraries screening centers.