期刊
JOURNAL OF BIOMOLECULAR SCREENING
卷 8, 期 5, 页码 489-499出版社
SAGE PUBLICATIONS INC
DOI: 10.1177/1087057103257309
关键词
gap junction; high-content screening; bioimaging; high-throughput screening; fluorescence microscopy-based imaging; automated fluorescence microscopy imaging
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.
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