Electron microscopy for ultrastructural analysis and protein localization in Saccharomyces cerevisiae
Authors:Andri Frankl, Muriel Mari and Fulvio Reggiori
Department of Cell Biology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
Keywords:
electron microscopy, electron tomography, immunolabeling, chemical fixation, cryo-immobilization, correlative light and electron microscopy, Saccharomyces cerevisiae
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The authors declare no conflict of interest.
Please cite this article as:
Andri Frankl, Muriel Mari and Fulvio Reggiori (2015). Electron microscopy for ultrastructural analysis and protein localization in Saccharomyces cerevisiae. Microbial Cell2(11): 412-428.
© 2015 Frankl et al. This is an open-access article released under the terms of the Creative Commons Attribution (CC BY) license, which allows the unrestricted use, distribution, and reproduction in any medium, provided the original author and source are acknowledged.
Abstract:
The yeast Saccharomyces cerevisiae is a key model system for studying of a multitude of cellular processes because of its amenability to genetics, molecular biology and biochemical procedures. Ultrastructural examinations of this organism, though, are traditionally difficult because of the presence of a thick cell wall and the high density of cytoplasmic proteins. A series of recent methodological and technical developments, however, has revived interest in morphological analyses of yeast (e.g. Mari et al., Traffic 2014; Kukulski et al., J Cell Biol 2011; Kuipers et al., Cell Tissue Res 2015). Here we present a review of established and new methods, from sample preparation to imaging, for the ultrastructural analysis of S. cerevisiae. We include information for the use of different fixation methods, embedding procedures, approaches for contrast enhancement, and sample visualization techniques, with references to successful examples. The goal of this review is to guide researchers that want to investigate a particular process at the ultrastructural level in yeast by aiding in the selection of the most appropriate approach to visualize a specific structure or subcellular compartment.
doi: 10.15698/mic2015.11.237
Volume 2, pp. 412 to 428, published 12/10/2015.