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 提高药物筛选项目的通量。这些检测检测何时 候选药物与目标受体结合，例如多巴胺， 神经降压素、阿片类药物和血清素受体，在 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、更通用且安全。 然而，现有的检测设备不足以满足 药物发现项目中的通量需求。制药公司 正在寻求吞吐量和灵敏度的改进。 我们建议以液晶光学和成像知识为基础 开发基于 CCD 的荧光微量滴定板读数器 偏振 (FP) 并显着提高灵敏度 与现有 FP 读板机相比的吞吐量。进步源自 新颖的光学系统，基于 CCD 的所有孔同时读取，以及 一种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源分析，显微镜- 基于实验来确认模型预测、面包板成像 实验和初步的光学设计。拟议的系统将 适用于任何板格式（96、384 或 1536 孔）和多标记 显示花期强度、资源能量转移的样品，和/或 极化。 拟议的商业应用： 我们的读板器将供以下药物发现部门使用 制药公司。较大的公司，其中有 至少 50 个，任何时候都有 20 到 50 个药物筛选项目正在进行 给定时间。大部分现有设备预计将被替换 当更高通量的仪器可用时。