title: High-throughput immunofluorescence microscopy using yeast spheroplast cell-based microarrays.
abstract: We have described a protocol for performing high-throughput immunofluorescence microscopy on microarrays of yeast cells. This approach employs immunostaining of spheroplasted yeast cells printed as high-density cell microarrays, followed by imaging using automated microscopy. A yeast spheroplast microarray can contain more than 5,000 printed spots, each containing cells from a given yeast strain, and is thus suitable for genome-wide screens focusing on single cell phenotypes, such as systematic localization or co-localization studies or genetic assays for genes affecting probed targets. We demonstrate the use of yeast spheroplast microarrays to probe microtubule and spindle defects across a collection of yeast strains harboring tetracycline-down-regulatable alleles of essential genes.
authors.affiliation: Department of Genetics, Yale University, New Haven, CT, USA. wei.niu08@gmail.com
authors.forename: Wei
authors.initials: W
authors.lastname: Niu
authors.forename: G Traver
authors.initials: GT
authors.lastname: Hart
authors.forename: Edward M
authors.initials: EM
authors.lastname: Marcotte
mesh_terms: D015536:Down-Regulation; D005800:Genes, Fungal; D057166:High-Throughput Screening Assays / Q000379:methods*; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism; D013104:Spheroplasts / Q000235:genetics*; D008941:Spindle Apparatus / Q000378:metabolism / Q000473:pathology; D013752:Tetracycline / Q000494:pharmacology; D046888:Tissue Array Analysis / Q000379:methods*; D015003:Yeasts / Q000235:genetics*
chemical_list: D013752:Tetracycline