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Identification of Gap Junction Blockers Using Automated Fluorescence Microscopy Imaging

Lead Discovery Technology, Lead Generation, julie.li{at}aventis.com

Yongping Yan

Lead Discovery Technology, Lead Generation

Elaine A. Powers

CNS Disease Group

Xiaoyou Ying

Bioimaging/Pathology/DSE, Lead Optimization, Aventis Pharmaceutical, Bridgewater, NJ

Khurram Janjua

Lead Discovery Technology, Lead Generation

Tina Garyantes

Lead Discovery Technology, Lead Generation

Bruce Baron

CNS Disease Group

Gap junctions coordinate electrical signals and facilitate metabolic synchronization between cells. In this study, the authors have developed a novel assay for the identification of gap junction blockers using fluorescence microscopy imaging-based high-content screening technology. In the assay, the communication between neighboring cells through gap junctions was measured by following the redistribution of a fluorescent marker. The movement of calcein dye from dye-loaded donor cells to dye-free acceptor cells through gap junctions overexpressed on cell surface membranes was monitored using automated fluorescence microscopy imaging in a high-throughput compatible format. The fluorescence imaging technology consisted of automated focusing, image acquisition, image processing, and data mining. The authors have successfully performed a high-throughput screening of a 486,000- compound program with this assay, and they were able to identify false positives without additional experiments. Selective and pharmacologically interesting compounds were identified for further optimization. (Journal of Biomolecular Screening 2003:489-499)

Key Words: gap junction • high-content screening • bioimaging • high-throughput screening • fluorescence microscopy-based imaging • automated fluorescence microscopy imaging

Journal of Biomolecular Screening, Vol. 8, No. 5, 489-499 (2003)
DOI: 10.1177/1087057103257309


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