Kinetic Imaging Plate Reader for Drug Discovery and Biology

项目总结 这项建议的目标是加强不同的生物医学科学家群体的能力， 西北大学为药物发现目的对活细胞进行光学分析，有针对性 药理学研究，监测细胞内第二信使(如cAMP，钙离子)，询问蛋白质- 蛋白质相互作用(使用FRET或BRET)，以及分析各种离子通道、转运蛋白的功能 和感受器。这一目标将通过购买和安装全光电磁动力系统来实现 由Wavefront Biosciences制造的成像平板阅读器。这台仪器将取代一台有10年历史的 西北大学高通量无功能荧光成像板阅读器(FLIPR-TETRA) 分析实验室(NUHTA)。全光EM是一款先进的动态成像印版阅读器，具有卓越的性能 特性和功能，包括1)高速动态成像荧光或发光信号 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 从紫外线到红外线的激发波长与最多5个发射滤光片相结合，覆盖 广泛的荧光和发光记者，3)具有近乎无失真图像的专有光学 质量与高速(高达100赫兹)电子倍增(EM)CCD摄像头相结合，4)速度快 波长切换，支持使用比率计量型染料、FRET或BRET的应用，5)同时多个 波长激发能力，支持光遗传和光药学应用，6)集成 高度灵活的液体搬运机器人，配有夹板器，以及强大、安全和用户友好的软件 数据采集、分析和管理。这些高级功能转化为高度灵活和 多功能成像平台，将为我们的设施提供实质性升级。 全景EM将由NU-HTA核心设施管理，该设施支持一系列技术 实现药物发现和生物学的高通量筛选。尽管NU-HTA将监督 仪器的管理，全景电磁将实际驻留在由 药理学部门，这将有助于随时访问本文件中列出的大多数主要用户和次要用户 申请。该仪器在药理学中的位置是与NU-HTA持续合作的一部分 这项计划旨在扩大西北大学芝加哥校区的服务。强有力的制度支持 将为该工具及其部署提供支持，并将由内部 咨询委员会。该仪器有望加强美国国立卫生研究院资助的几项研究的成功 项目，并为积极和未来的研究提供一个新的平台。

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.