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。研制了多光谱拉曼系统（MRS）作为uHTS的光学读出。目的2：使用分子哨兵评估uHTS-MRS用于高度平行的配体/靶结合检测。 目标3：开发和集成自动化系统，用于检测基于SERS的纳米探针的光信号。 目的4：评估CMOS-uHTS系统在细胞模型中用于小分子或mRNA探针的高通量筛选。 我们的系统将提供一个新的层面，在标签复用表征大量的样品在基于细胞的高通量筛选药物发现。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。提出的系统的独特功能包括：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“标记多重”检测（即同时使用许多拉曼染料标记探针的能力）（极窄的拉曼带允许Sers用作多重技术）。 将测试该系统以筛选上调或下调各种mRNA家族表达的小分子、合成化学品和天然产物文库。将鉴定能够调节mRNA基因表达水平的活性化合物，然后进行额外的测定。基于SERS的uHTS仪器将提供一种工具，该工具具有比目前可用的系统更快和更有效的潜力，并且能够提供具有更高特异性水平的高灵敏度和多路复用能力，以筛选合成化学和天然产物文库，例如将在NIH赞助的分子文库筛选中心注册和安置的文库。

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.