Building cell structures in three dimensions: Electron tomography methods for budding yeast

Eileen T. O’Toole, Thomas H. Giddings, Mark Winey

Research output: Contribution to journalArticle

Abstract

Saccharomyces cerevisiae has been an important model system for numerous cellular, genetic, and molecular studies. However, this small eukaryote presents a challenge for imaging at the electron microscope level. Preparation of yeast using high-pressure freezing followed by freeze-substitution (HPF/FS) results in excellent preservation of cell structure in these difficult-to-fix samples. In particular, cells prepared by HPF/FS can be used for 3D electron tomography (ET) studies where optimum cell preservation is critical. Here, we discuss the advantages of using HPF/FS for ET and show examples of the utility of this method for building yeast cell structures in three dimensions.

Original languageEnglish (US)
Pages (from-to)183-187
Number of pages5
JournalCold Spring Harbor Protocols
Volume2017
Issue number3
DOIs
StatePublished - Mar 1 2017
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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