CCD-BASED FLUORESCENCE POLARIZATION ASSAY PLATE READER

基于 CCD 的荧光偏振分析读板机

• 批准号: 2648268
• 负责人: CLIFFORD C HOYT
• 金额: $ 9.99万
• 依托单位: CAMBRIDGE RESEARCH AND INSTRUMENTATION
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-15 至 1999-03-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2648268
• 关键词: biomedical equipment development; charge coupled device camera; drug screening /evaluation; fluorescence polarization; image processing; liquid crystal; method development

Fluorescence polarization-based assays hold tremendous promise for increasing throughput of drug screening programs. These assays detect when candidate drugs bind to target receptors, such as the dopamine, neurotensin, opioid, and serotonin receptors, which play key roles in the central nervous system. Compared to conventional radio-isotope assays, fluorescence assays are faster, simpler, and more versatile and safe. However, existing equipment for performing assays is in sufficient to meet the throughput needs in drug discovery programs. Pharmaceutical companies are looking for improvements in throughput and sensitivity. We propose to build on our knowledge of liquid crystal optics and imaging to develop a CCD-based microtitre plate reader that uses fluorescence polarization (FP) and that offers significant improvements in sensitivity and throughput compared to existing FP plate readers. Advances derive from a novel optical system, CCD-based simultaneous reading of all wells, and a novel background compensation scheme. Proposed Phase I work includes an analysis of sources of noise and error in FP measurements, microscope- based experiments to confirm model predictions, breadboard imaging experiments, and a preliminary optical design. The proposed system will work with any plate format (96, 384, or 1536 wells) and with multi-labeled samples exhibiting florescence intensity, resource energy transfer, and/or polarization. PROPOSED COMMERCIAL APPLICATION: Our plate reader would be used by drug discovery departments of pharmaceutical companies. The larger companies, of which there are at least 50, have between 20 and 50 drug screening programs going on at any given time. Most of the existing equipment is expected to be replaced by higher throughput instruments, when they become available.

基于荧光偏振的测定具有巨大的前景， 增加药物筛选程序的吞吐量。这些检测试剂盒检测何时 候选药物与靶受体结合，如多巴胺， 神经降压素、阿片样物质和5-羟色胺受体，它们在神经系统中起着关键作用。 中枢神经系统与传统的放射性同位素测定相比， 荧光测定更快、更简单、更通用和更安全。 然而，用于进行测定的现有设备不足以满足 药物发现项目中的产量需求。制药公司 正在寻求提高吞吐量和灵敏度。 我们建议建立在我们的知识液晶光学和成像 开发一种基于CCD的微量滴定板读数器， 偏振（FP），并提供显着改善灵敏度 和通量。进步源于 新颖的光学系统，基于CCD的所有威尔斯的同时阅读，和 新颖的背景补偿方案。拟议的第一阶段工作包括 FP测量中的噪声和误差来源分析，显微镜- 基于实验来确认模型预测，实验板成像 实验和初步的光学设计。拟设系统可 使用任何板格式（96、384或1536威尔斯孔）和多标记 显示出荧光强度、资源能量转移和/或 极化 拟定商业应用： 我们的平板阅读器将被用于药物研发部门， 制药公司。较大的公司，其中有在 至少有50个，有20到50个药物筛选项目正在进行， 给你时间预计大部分现有设备将由 更高通量的仪器，当它们变得可用时。