Abstract
Budding yeast Saccharomyces cerevisiae represents a widely used model organism for the study of mitochondrial biogenesis and architecture. Electron microscopy is an essential tool in the analysis of cellular ultrastructure and the precise localization of proteins to organellar subcompartments. We provide here detailed protocols for the analysis of yeast mitochondria by transmission electron microscopy: (1) chemical fixation and Epon embedding of yeast cells and isolated mitochondria, and (2) cryosectioning and immunolabeling of yeast cells and isolated mitochondria according to the Tokuyasu method.
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Acknowledgments
We thank Fulvio Reggiori and Muriel Mari (University Medical Centre Utrecht, The Netherlands) for advice and excellent training in cryo-sectioning, Rita Grotjahn for technical assistance, and Anja Heublein for photography. This work was supported by the Elitenetzwerk Bayern and a Max-Planck-Fellowship to A.-K.U. and grants of the Deutsche Forschungsgemeinschaft to B.W.
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Unger, AK., Geimer, S., Harner, M., Neupert, W., Westermann, B. (2017). Analysis of Yeast Mitochondria by Electron Microscopy. In: Mokranjac, D., Perocchi, F. (eds) Mitochondria. Methods in Molecular Biology, vol 1567. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6824-4_18
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DOI: https://doi.org/10.1007/978-1-4939-6824-4_18
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