Work

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

Year: 2010

Type: book-chapter

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.

Source: Methods in molecular biology

DOI: 10.1007/978-1-61737-970-3_7

Authors: Wei Niu, G. Traver Hart, Edward M. Marcotte

Institutions: Yale University, The University of Texas at Austin

Language: English

Topic: Cell Image Analysis Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funder: National Institute of General Medical Sciences

Award: Proteomics of widespread, reversible protein assemblies in aging and aggregation