Kinetic Imaging Plate Reader for Drug Discovery and Biology

点击翻译按钮获取中文摘要

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.