Microwave-assisted preparation of yeast cells for ultrastructural analysis by electron microscopy
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 desc...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Shared Science Publishers OG
2024-11-01
|
| Series: | Microbial Cell |
| Subjects: | |
| Online Access: | http://microbialcell.com/researcharticles/2024a-mayer-microbial-cell |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850053040993730560 |
|---|---|
| author | Moritz Mayer Christina Schug Stefan Geimer Till Klecker Benedikt Westermann |
| author_facet | Moritz Mayer Christina Schug Stefan Geimer Till Klecker Benedikt Westermann |
| author_sort | Moritz Mayer |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-be0bbd312f754c048486cfbca1f786a3 |
| institution | DOAJ |
| issn | 2311-2638 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Shared Science Publishers OG |
| record_format | Article |
| series | Microbial Cell |
| spelling | doaj-art-be0bbd312f754c048486cfbca1f786a32025-08-20T02:52:38ZengShared Science Publishers OGMicrobial Cell2311-26382024-11-011137838610.15698/mic2024.11.840Microwave-assisted preparation of yeast cells for ultrastructural analysis by electron microscopyMoritz Mayer0https://orcid.org/0009-0001-1283-5115Christina Schug1https://orcid.org/0009-0005-7270-4984Stefan Geimer2https://orcid.org/0000-0001-6095-9721Till Klecker3https://orcid.org/0000-0003-0149-042XBenedikt Westermann4https://orcid.org/0000-0002-2991-1604Zellbiologie und Elektronenmikroskopie, Universität Bayreuth, Bayreuth, 95440, GermanyZellbiologie und Elektronenmikroskopie, Universität Bayreuth, Bayreuth, 95440, GermanyZellbiologie und Elektronenmikroskopie, Universität Bayreuth, Bayreuth, 95440, GermanyZellbiologie und Elektronenmikroskopie, Universität Bayreuth, Bayreuth, 95440, GermanyZellbiologie und Elektronenmikroskopie, Universität Bayreuth, Bayreuth, 95440, GermanyBudding 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.http://microbialcell.com/researcharticles/2024a-mayer-microbial-cellmicrowave fixationmitochondriaorganelle architecturepotassium permanganate fixationtransmission electron microscopysaccharomyces cerevisiae |
| spellingShingle | Moritz Mayer Christina Schug Stefan Geimer Till Klecker Benedikt Westermann Microwave-assisted preparation of yeast cells for ultrastructural analysis by electron microscopy Microbial Cell microwave fixation mitochondria organelle architecture potassium permanganate fixation transmission electron microscopy saccharomyces cerevisiae |
| title | Microwave-assisted preparation of yeast cells for ultrastructural analysis by electron microscopy |
| title_full | Microwave-assisted preparation of yeast cells for ultrastructural analysis by electron microscopy |
| title_fullStr | Microwave-assisted preparation of yeast cells for ultrastructural analysis by electron microscopy |
| title_full_unstemmed | Microwave-assisted preparation of yeast cells for ultrastructural analysis by electron microscopy |
| title_short | Microwave-assisted preparation of yeast cells for ultrastructural analysis by electron microscopy |
| title_sort | microwave assisted preparation of yeast cells for ultrastructural analysis by electron microscopy |
| topic | microwave fixation mitochondria organelle architecture potassium permanganate fixation transmission electron microscopy saccharomyces cerevisiae |
| url | http://microbialcell.com/researcharticles/2024a-mayer-microbial-cell |
| work_keys_str_mv | AT moritzmayer microwaveassistedpreparationofyeastcellsforultrastructuralanalysisbyelectronmicroscopy AT christinaschug microwaveassistedpreparationofyeastcellsforultrastructuralanalysisbyelectronmicroscopy AT stefangeimer microwaveassistedpreparationofyeastcellsforultrastructuralanalysisbyelectronmicroscopy AT tillklecker microwaveassistedpreparationofyeastcellsforultrastructuralanalysisbyelectronmicroscopy AT benediktwestermann microwaveassistedpreparationofyeastcellsforultrastructuralanalysisbyelectronmicroscopy |