Document Detail


High-throughput immunofluorescence microscopy using yeast spheroplast cell-based microarrays.
MedLine Citation:
PMID:  21104056     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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:
Wei Niu; G Traver Hart; Edward M Marcotte
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  706     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2011  
Date Detail:
Created Date:  2010-11-24     Completed Date:  2011-03-22     Revised Date:  2013-07-03    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  83-95     Citation Subset:  IM    
Affiliation:
Department of Genetics, Yale University, New Haven, CT, USA. wei.niu08@gmail.com
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MeSH Terms
Descriptor/Qualifier:
Down-Regulation
Genes, Fungal
High-Throughput Screening Assays / methods*
Microscopy, Fluorescence
Microtubules / metabolism
Mitotic Spindle Apparatus / metabolism,  pathology
Spheroplasts / genetics*
Tetracycline / pharmacology
Tissue Array Analysis / methods*
Yeasts / genetics*
Grant Support
ID/Acronym/Agency:
R01 GM076536/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
60-54-8/Tetracycline
Comments/Corrections

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