This proposal seeks to develop a unique high throughput screening (HTS) system composed of several integrated existing and novel automated components that will be uniquely capable of screening entire protein target families simultaneously. The following advantages will be realized due to its maximized efficiencies: i) improved drug selectivity and a reduction in drug side effects, and ii) greater than ten fold reduction in the cost of screening and drug candidate optimization. Additionally, novel instrumentation and methodologies will become available to the biomedical research community for academic and industrial use, including a high speed, six laser flow cytometer developed through DakoCytomation the (BRP partner) and an automated tissue culture system developed through The Automation Partnership. The Specific Aims are i) to assist in development of and procure an automated tissue culture system capable of handling over 300 microwell plates and nearly 2,000 distinct cell lines, ii) to expand the capabilities of our automated sampling system for flow cytometry (FCM) for greater speed and to enable unique analysis and sorting modes, iii) to develop at least 150 GPCR target cell lines through novel transfection and expression techniques and cellular evolution techniques, iv) to develop a single cell HTS screening platform based on six-laser FCM, multiparametric detection technology and an automated cell preparation system, v) to perform a focused library test screen of 10,000 compounds against the 150 GPCR targets, and vi) to develop the database and bioinformatics tools to integrate and help interpret the data. When fully integrated, tested and validated the system will be applicable to many target families of relevance to biomedical and pharmaceutical research, such as other receptor classes, and will have a broad range of drug discovery applications. [unreadable] [unreadable]