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Hidalgo, I.H.* ; Fleming, T.* ; Eckstein, V.* ; Herzig, S. ; Nawroth, P.P.* ; Tyedmers, J.*

Characterization of aggregate load and pattern in living yeast cells by flow cytometry.

BioTechniques 61, 137-148 (2016)
Publ. Version/Full Text Research data DOI
Open Access Gold
Protein aggregation is both a hallmark of and a driving force for a number of diseases. It is therefore important to identify the nature of these aggregates and the mechanism(s) by which the cell counteracts their detrimental properties. Currently, the study of aggregation in vivo is performed primarily using fluorescently tagged versions of proteins and analyzing the aggregates by fluorescence microscopy. While this strategy is considered the gold standard, it has several limitations, particularly with respect to its suitability for high-throughput screening (HTS). Here, using a GFP fusion of the well-characterized yeast prion amyloid protein [PSI+], we demonstrate that flow cytometry, which utilizes the same physical principles as fluorescence microscopy, can be used to determine the aggregate load and pattern in live and fixed yeast cells. Furthermore, our approach can easily be applied to high-throughput analyses such as screenings with a yeast deletion library.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Amyloid Aggregates ; Flow Cytometry ; Fluorescence Activated Cell Sorting (facs) ; High-throughput Screening (hts) ; Protein Aggregation ; Saccharomyces Cerevisiae ; Yeast Prion; Quality-control; Prion Fragmentation; Alpha-synuclein; Psi+ Prion; Proteins; Inheritance; Hsp104; Hsp70; Segregation; Elimination
ISSN (print) / ISBN 0736-6205
e-ISSN 1940-9818
Journal BioTechniques / Protocol Guide
Quellenangaben Volume: 61, Issue: 3, Pages: 137-148 Article Number: , Supplement: ,
Publisher Informa Life Sciences Publ.
Publishing Place New York, NY
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Diabetes and Cancer (IDC)