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

描述(申请人提供)：本研究的目标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。本项目的具体目标是：目标1.研制一种多光谱拉曼系统(MRS)作为超高温超导的光学读出系统。目的2：利用分子哨兵技术对uHTS-MRS进行高度平行的配体/靶结合检测。目的3：开发和集成用于检测基于SERS的纳米探针的光信号的自动化系统。目的：在细胞模型中评价高通量筛选小分子或信使核糖核酸探针的CMOS-uHTS系统。我们的系统将为基于细胞的高通量药物筛选中表征大量样本的标记多路复用提供一个新的维度。一种新的基于SERS的分子前哨检测方案将被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效应使常规拉曼信号的灵敏度提高了106-1015倍，SERS-MS纳米探针对光漂白(通常与荧光探针结合)不敏感。超高通量“标记多路传输”检测(即同时使用多个拉曼染料标记探针的能力)(极窄的拉曼频带允许将SERS用作多路传输技术)。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确定，随后将进行额外的分析。基于SERS的uHTS仪器将提供一种可能比目前可用的系统更快、更高效的工具，能够提供极高的灵敏度和更高水平的特异度，并具有多路复用能力来筛选合成化学和天然产品库，例如将注册并存放在NIH赞助的分子库筛选中心的库。

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.