Cell-Based Microarrays pp 83-95

Part of the Methods in Molecular Biology book series (MIMB, volume 706)

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

Protocol

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 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of GeneticsYale UniversityNew HavenUSA
  2. 2.Department of Chemistry and BiochemistryCenter for Systems and Synthetic Biology, Institute for Cellular and Molecular Biology, University of Texas at AustinAustinUSA

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