High-Throughput Imaging of Arrays of Fluorescently Tagged Yeast Mutant Strains

We describe a protocol for live-cell high-throughput (HTP) screening of yeast mutant strains carrying fluorescent protein markers for subcellular compartments of choice using automated confocal microscopy. This procedure, which combines HTP genetics and microscopy, results in the acquisition of thousands of images that can be analyzed in a systematic and quantitative way to identify morphology defects in the tagged subcellular compartments. This HTP protocol is readily adapted for screening any combination of markers and can be expanded to different growth conditions or higher order mutant genetic backgrounds.