In situ high-resolution cryo-EM reconstructions from CEMOVIS.
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Description
Johannes Elferich and Marek Kaminek contributed equally to this work.
BORIS DOI
Date of Publication
July 1, 2025
Publication Type
Article
Division/Institute
Subject(s)
Series
IUCrJ
ISSN or ISBN (if monograph)
2052-2525
Publisher
International Union of Crystallography
Language
English
Publisher DOI
PubMed ID
40553549
Description
Cryo-electron microscopy can be used to image cells and tissue at high resolution. To ensure electron transparency, the sample thickness must not exceed 500 nm. Focused-ion-beam (FIB) milling has become the standard method for preparing thin samples (lamellae); however, the material removed by the milling process is lost, the imageable area is usually limited to a few square micrometres and the surface layers sustain damage from the ion beam. We have examined cryo-electron microscopy of vitreous sections (CEMOVIS), a technique based on cutting thin sections with a knife, as an alternative to FIB milling. Vitreous sections also sustain damage, including compression, shearing and cracks. However, samples can be sectioned in series, producing many orders of magnitude more imageable area compared to lamellae, making CEMOVIS an alternative to FIB milling with distinct advantages. Using two-dimensional template matching on images of vitreous sections of Saccharomyces cerevisiae cells, we reconstructed the 60S ribosomal subunit at near-atomic resolution, demonstrating that, in many regions of the sections, the molecular structure of these subunits is largely intact, comparable to FIB-milled lamellae.
File(s)
File | File Type | Format | Size | License | Publisher/Copright statement | Content | |
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eh5022.pdf | text | Adobe PDF | 9.01 MB | Attribution (CC BY 4.0) | published |