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

Phenomics approaches to understand genetic networks and gene function in yeast

“…In addition to facilitating rapid construction of double mutant yeast arrays (see above), the SGA method can also be used to introduce fluorescent markers of any subcellular compartment of interest arrayed collections of yeast mutants (Figure 1). Additional markers to provide spatial context, such as nuclear and cytoplasmic markers, can also be simultaneously introduced enabling cell segmentation and automated image analysis [46]. This general approach has been applied over the last decade or so, using various imaging systems and image analysis protocols, to discover phenotypes caused by single-gene perturbations for a spectrum of subcellular structures and processes including: (1) morphology of the mitotic spindle [47]; (2) determinants of nuclear shape [48,49]; (3) nucleolar size regulation [50]; (4) regulation of trafficking at endosomes [51]; (5) formation of DNA damage foci [52] and; (6) endocytosis compartment morphology and regulators of endocytosis [53][54][55].…”

“…In addition to facilitating rapid construction of double mutant yeast arrays (see above), the SGA method can also be used to introduce fluorescent markers of any subcellular compartment of interest into arrayed collections of yeast mutants ( Figure 1 ). Additional markers to provide spatial context, such as nuclear and cytoplasmic markers, can also be simultaneously introduced enabling cell segmentation and automated image analysis [ 46 ].…”