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

Itahisa Hernández Hidalgo, Thomas Fleming, Volker Eckstein, Stephan Herzig, Peter P. Nawroth, Jens Tyedmers

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)137-148
Number of pages12
JournalBioTechniques
Volume61
Issue number3
DOIs
StatePublished - Sep 2016
Externally publishedYes

Keywords

  • Amyloid aggregates
  • Flow cytometry
  • Fluorescence activated cell sorting (FACS)
  • High-throughput screening (HTS)
  • Protein aggregation
  • Saccharomyces cerevisiae
  • Yeast prion

Fingerprint

Dive into the research topics of 'Characterization of aggregate load and pattern in living yeast cells by flow cytometry'. Together they form a unique fingerprint.

Cite this