Kinetic Imaging Plate Reader for Drug Discovery and Biology

项目摘要 这项建议的目的是提高生物医学科学家的能力， 西北大学对活细胞进行光学分析，用于药物发现，靶向 药理学研究，监测细胞内第二信使（例如，cAMP，Ca 2+），询问蛋白- 蛋白质相互作用（使用FRET或BRET），并测定各种离子通道，转运蛋白 和受体。这一目标将通过购买和安装Panoptic EM动能来实现 由WaveFront Biosciences制造的成像板读取器。这台仪器将取代10年前的 非功能性荧光成像板阅读器（FLIPR-tetra）在西北大学高中 分析实验室（NU-HTA）。Panoptic EM是一款先进的动态成像酶标仪， 特征和功能包括：1）来自以下物质的荧光或发光信号的高速动力学成像： 96、384或1536孔板，2）发射7个离散窄带的长寿命固态激光光源， 激发波长范围从紫外线到红外线，与多达5个发射滤光片耦合， 广泛的荧光和发光报告，3）专有光学与近失真的图像 高质量加上高速（高达100 Hz）电子倍增（EM）CCD相机，4）快速 波长切换以实现使用比率染料、FRET或BRET的应用，5）同时多个 波长激发能力，以实现光遗传学和光药理学应用，6）集成 具有平板夹持器的高度灵活的液体处理机器人，以及7）强大，安全和用户友好的软件， 数据采集、分析和管理。这些先进的功能转化为高度灵活， 多功能成像平台，将为我们的设施提供实质性升级。 Panoptic EM将由NU-HTA核心设施管理，该设施支持一系列技术 从而能够进行用于药物发现和生物学的高通量筛选。尽管NU-HTA将监督 为了管理仪器，Panoptic EM将物理地驻留在由 药理学系，这将有助于随时访问大多数主要和次要用户命名在此 应用程序.该仪器在药理学中的放置是与NU-HTA正在进行的合作的一部分 旨在扩大西北大学芝加哥校区的服务。有力制度支撑 将提供该工具及其部署的内部监督，并将通过一个内部监督机构进行有效监督。 咨询委员会。该仪器有望提高NIH资助的几项研究的成功率 项目，并为积极和未来的研究提供了一个新的平台。

PROJECT SUMMARY The objective of this proposal is to enhance the capability of a diverse group of biomedical scientists at Northwestern University to perform optical assays on live cells for the purposes of drug discovery, targeted pharmacological studies, monitoring intracellular second messengers (e.g., cAMP, Ca2+), interrogating protein- protein interactions (using FRET or BRET), and assaying the function of a variety of ion channels, transporters and receptors. This objective will be achieved by the purchase and installation of a Panoptic EM kinetic imaging plate reader manufactured by WaveFront Biosciences. This instrument will replace a 10-year old nonfunctional fluorescence imaging plate reader (FLIPR-tetra) in the Northwestern University High Throughput Analysis Laboratory (NU-HTA). The Panoptic EM is an advanced kinetic imaging plate reader with exceptional features and functionalities including 1) high speed kinetic imaging of fluorescent or luminescent signals from 96, 384, or 1536-well plates, 2) a long lifetime solid state laser light source emitting 7 discrete narrow band excitation wavelengths ranging from ultraviolet to infrared coupled with up to 5 emission filters that covers a wide range of fluorescent and luminescent reporters, 3) proprietary optics with near distortion-free image quality coupled with a high speed (up to 100 Hz) electron multiplying (EM) CCD camera, 4) rapid wavelength switching to enable applications using ratiometric dyes, FRET or BRET, 5) simultaneous multiple wavelength excitation capability to enable optogenetic and photo-pharmacological applications, 6) integrated highly flexible liquid handing robot with plate gripper, and 7) powerful, secure and user-friendly software for data acquisition, analysis and management. These advanced features translate into a highly flexible and versatile imaging platform that will provide a substantial upgrade for our facility. The Panoptic EM will be managed by the NU-HTA core facility, which supports a range of technologies enabling high throughput screening for drug discovery and biology. Although NU-HTA will oversee management of the instrument, the Panoptic EM will physically reside within space provided by the Department of Pharmacology, which will facilitate ready access to most Major and Minor users named in this application. The placement of the instrument in Pharmacology is part of an ongoing collaboration with NU-HTA that seeks to expand services on the Chicago campus of Northwestern University. Strong institutional support for the instrument and its deployment will be provided, and effective oversight will be achieved by an internal advisory committee. The instrument is expected to enhance the success of several NIH-funded research projects and provide a new platform for active and future research.