2022
DOI: 10.20944/preprints202203.0146.v1
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Expansion Microscopy on <em>Saccharomyces cerevisiae</em>

Abstract: The unicellular eukaryote S. cerevisiae is an invaluable resource for the study of basic eukaryotic cellular and molecular processes. However, due to its small size compared to other eukaryotic organisms the study of subcellular structures is challenging. Expansion microscopy (ExM) holds great potential to study the intracellular architecture of yeast, especially when paired with pan-labelling techniques visualising the full protein content inside cells. ExM allows to increase imaging resolution by physically … Show more

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Cited by 6 publications
(8 citation statements)
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“…However, such approaches include a full enzymatic treatment that digests the proteome, and more importantly, pre-expansion labeling which retains the linkage error (Zwettler et al, 2020). Post-labeling ExM protocol on as S. Cerevisiae has recently been reported (Korovesi et al, 2022); however, the resolution power of this method on different organelles, its compatibility with antibodies, and its feasibility in other yeast models remain to be assessed. Here, we develop a robust U-ExM protocol for budding and fission yeast, keeping the proteome intact and labelling post-expansion to reduce linkage errors (Hamel and Guichard, 2021).…”
Section: Introductionmentioning
confidence: 99%
“…However, such approaches include a full enzymatic treatment that digests the proteome, and more importantly, pre-expansion labeling which retains the linkage error (Zwettler et al, 2020). Post-labeling ExM protocol on as S. Cerevisiae has recently been reported (Korovesi et al, 2022); however, the resolution power of this method on different organelles, its compatibility with antibodies, and its feasibility in other yeast models remain to be assessed. Here, we develop a robust U-ExM protocol for budding and fission yeast, keeping the proteome intact and labelling post-expansion to reduce linkage errors (Hamel and Guichard, 2021).…”
Section: Introductionmentioning
confidence: 99%
“…141 This approach was replicated almost identically by another group not much later. 142 A specific Chitinase seems unnecessary to obtain full 4-fold homogeneous expansion of yeast cells, likely owing to its low quantity in yeast cell walls.…”
Section: ■ Preparing Diverse Organisms and Sample Types For Expansion...mentioning
confidence: 99%
“…Lessons from Cell Wall Homogenization: Adapting Existing Protocols. Of note, both the authors of μExM 37 and the groups that achieved expansion of Saccharomyces cerevisiae 39,142 were adapting protocols originally used for other methods relating to their model organisms. Bacteria are lysed with lysozyme and mutanolysin to obtain spheroblasts for transformations or other processes, or for DNA preparations� just as the protein digestion approach in classical expansion microscopy 1 was adapted from DNA-extraction protocols.…”
Section: ■ Preparing Diverse Organisms and Sample Types For Expansion...mentioning
confidence: 99%
See 1 more Smart Citation
“…While traditionally considered suboptimal for the imaging of intracellular features, superresolution techniques were recently developed (Chen et al, 2021;Hinterndorfer et al, 2022;Korovesi et al, 2022), and used (Dey et al, 2020) in yeast to study cellular processes at higher resolution, and new tools have been provided for the imaging of several subcellular structures (Akhuli et al, 2022;Zhu et al, 2019). A large number of telomere-to-telomere assemblies recently revealed the landscape of structural variants of S. cerevisiae natural isolates (S. O'Donnell et al, 2022).…”
Section: Conclusion and Future Perspectivesmentioning
confidence: 99%