Multipixel Hybrid Photon-Counting Detector for High-Throughput Single-Molecule As

描述(由申请人提供)：该项目的长期目标是为生物医学研究团体提供一种用于高通量单分子荧光分析的新设备。这些方法的应用领域从基础研究到高通量诊断和药物筛选。这项研究计划的目标是开发一种用于多路高通量单分子分析的集成系统。该系统将基于滨松光电株式会社最近开发的一种新型的单光子计数能力的多通道混合光电探测器(HPD)。我们将把这种探测器集成到多激发/多检测点共聚焦显微镜中，并在实际的单分子分析中展示其能力。我们期望这种探测器不仅应用于单分子方法的基础研究应用，还将应用于高通量单分子诊断和药物筛选等新兴领域。本研究方案的具体目标是：(1)为滨松公司开发的多通道混合探测器开发电子接口和数据采集系统(包括软件)。(2)将该系统集成到多路单分子荧光激发/检测共聚焦显微镜中。(3)通过对与生物相关的模型系统进行多路复用荧光相关光谱(FCS)测量以及使用交替激光激发(Alex)方案进行多路复用单分子FRET测量，来展示该系统用于多路复用、高通量单分子分析的能力。

DESCRIPTION (provided by applicant): The long-term objective of this project is to provide the biomedical research community with a new device for high-throughput single-molecule fluorescence assays. The domains of applications of these methods range from basic research to high-throughput diagnostics and drug screening. The goal of this research proposal is to develop an integrated system for multiplexed, high-throughput single-molecule assays. This system will be based on a new type of single-photon counting capable mutichannel hybrid photodetector (HPD) recently developed by Hamamatsu Photonics Corporation. We will incorporate this detector into a multiple excitation/multiple detection spot confocal microscope and demonstrate its capabilities in actual single-molecule assays. We expect this detector to find applications not only in fundamental research applications using single-molecule methods, but also in the emerging fields of high-throughput single-molecule diagnostics and drug screening. The specific aims of this research proposal are: (1) to develop an electronic interface and data acquisition system (including software) for the multichannel hybrid detector developed by Hamamatsu. (2) to integrate this system into a multiplexed single-molecule fluorescence excitation/detection confocal microscope. (3) to demonstrate the capabilities of this system for multiplexed, high-throughput single- molecule assays, by performing multiplexed fluorescence correlation spectroscopy (FCS) measurements on model systems of biological relevance as well as multiplexed single-molecule FRET measurements using an alternate laser excitation (ALEX) scheme.