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.
CITATION STYLE
Niu, W., Traver Hart, G., & Marcotte, E. M. (2011). High-throughput immunofluorescence microscopy using yeast spheroplast cell-based microarrays. In Methods in Molecular Biology (Vol. 706, pp. 83–95). Humana Press Inc. https://doi.org/10.1007/978-1-61737-970-3_7
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