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TitleIn situ high-resolution cryo-EM reconstructions from CEMOVIS.
Journal, issue, pagesIucrj, Year 2025
Publish dateJun 6, 2025
AuthorsJohannes Elferich / Marek Kaminek / Lingli Kong / Adolfo Odriozola / Wanda Kukulski / Benoît Zuber / Nikolaus Grigorieff /
PubMed AbstractCryo-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 ...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.
External linksIucrj / PubMed:40553549 / PubMed Central
MethodsEM (single particle)
Resolution3.5 Å
Structure data

EMDB-71068: Reconstruction of the yeast 60S ribosomal subunit from CEMOVIS sections
Method: EM (single particle) / Resolution: 3.5 Å

Source
  • Saccharomyces cerevisiae (brewer's yeast)

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