Protocol

Cell-Based Microarrays

Volume 706 of the series Methods in Molecular Biology pp 83-95

Date:

High-Throughput Immunofluorescence Microscopy Using Yeast Spheroplast Cell-Based Microarrays

  • Wei NiuAffiliated withDepartment of Genetics, Yale University
  • , G. Traver HartAffiliated withDepartment of Chemistry and Biochemistry, Center for Systems and Synthetic Biology, Institute for Cellular and Molecular Biology, University of Texas at Austin
  • , Edward M. MarcotteAffiliated withDepartment of Chemistry and Biochemistry, Center for Systems and Synthetic Biology, Institute for Cellular and Molecular Biology, University of Texas at Austin Email author 

* Final gross prices may vary according to local VAT.

Get Access

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.

Key words

Yeast immunofluorescence high-throughput microscopy cell microarrays microtubule