Cryopreparation and electron tomography of yeast cells

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

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Three-dimensional imaging of cells using electron tomography enables analysis of cell structure at unprecedented resolution. The preparation of cells for tomography using rapid freezing followed by freeze-substitution is an essential first step to ensure the optimal preservation of the cell structure for 3D studies. This protocol outlines a method for obtaining well-preserved cells using high-pressure freezing followed by freeze-substitution. We have found that this method is particularly well suited for electron tomography studies and has the added bonus of preserving antigenicity for immuno-electron microscopy. The steps involved in imaging cells and performing tomographic analysis of cellular structures are also outlined.

Original languageEnglish (US)
Pages (from-to)212-216
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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