Reference: O'Toole ET, et al. (2017) Building Cell Structures in Three Dimensions: Electron Tomography Methods for Budding Yeast. Cold Spring Harb Protoc 2017(3)

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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.

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Journal Article
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O'Toole ET, Giddings TH, Winey M
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