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High-content screening of yeast mutant librariesHigh-content screening assays have enabled the efficient analysis of cellular events in a large number of experimental samples and with high throughput. Screening approaches based on light microscopic techniques have been applied e.g. to characterize morphological phenotypes of cells caused by mutations or by treatment of cells with chemical inhibitors or drugs. In addition, screening assays using fluorescence microscopy particularly in combination with green-fluorescent-protein (GFP) technology have provided a more global view of the spatial and temporal behavior of subcellular structures in the context of the living cell.
We have recently developed a high-resolution cell-based assay in intact yeast cells to systematically screen yeast mutant strain libraries for abnormal morphology and distribution of fluorescently labeled subcellular structures. The method was validated in a pilot screen assaying abnormal morphology and distribution of fluorescently labeled peroxisomes of yeast. The modular assay enables reliable imaging-based analysis of fluorescently labeled subcellular structures of entire yeast strain collections (1,2).
Flow chart of the established cell-based assay in intact yeast cells
for high-content mutant and drug screens.
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