Microwave-assisted preparation of yeast cells for ultrastructural analysis by electron microscopy

Authors:

Moritz Mayera, Christina Schuga, Stefan Geimer, Till Klecker and Benedikt Westermann

doi: 10.15698/mic2024.11.840
Volume 11, pp. 378 to 386, published 18/11/2024.

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Affiliations:

Zellbiologie und Elektronenmikroskopie, Universität Bayreuth, 95440 Bayreuth, Germany.

a These authors contributed equally to this work.

Keywords: 

microwave fixation, mitochondria, organelle architecture, potassium permanganate fixation, transmission electron microscopy, Saccharomyces cerevisiae.

Corresponding Author(s):

Till Klecker, Zellbiologie und Elektronenmikroskopie, Universität Bayreuth, 95440 Bayreuth, Germany; Phone: +49-921 55 4311; till.klecker@uni-bayreuth.de Benedikt Westermann, Zellbiologie und Elektronenmikroskopie, Universität Bayreuth, 95440 Bayreuth, Germany; Phone: +49-921 55 4300; benedikt.westermann@uni-bayreuth.de

Conflict of interest statement:

The authors declare that there are no conflicts of interest.

Please cite this article as:

Moritz Mayer, Christina Schug, Stefan Geimer, Till Klecker and Benedikt Westermann (2024). Microwave-assisted preparation of yeast cells for ultrastructural analysis by electron microscopy. Microbial Cell 11: 378-386. doi: 10.15698/mic2024.11.840

© 2024 Mayer 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:

Budding yeast Saccharomyces cerevisiae is widely used as a model organism to study the biogenesis and architecture of organellar membranes, which can be visualized by transmission electron microscopy (TEM). Preparation of yeast cells for TEM can be quite challenging and time-consuming. Here, we describe an optimized protocol for conventional fixation of yeast cells with potassium permanganate combined with cell wall permeabilization with sodium metaperiodate and embedding in Epon. We have replaced time-consuming incubation steps by short treatments with microwaves and developed a microwave-assisted permanganate fixation and Epon embedding protocol that reduces the time required for sample preparation to one working day. We expect that these protocols will be useful for routine analysis of membrane ultrastructure in yeast.