In situ high-resolution cryo-EM reconstructions from CEMOVIS
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 m...
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International Union of Crystallography
2025-07-01
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| Series: | IUCrJ |
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| Online Access: | https://journals.iucr.org/paper?S2052252525005196 |
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| author | Johannes Elferich Marek Kaminek Lingli Kong Adolfo Odriozola Wanda Kukulski Benoît Zuber Nikolaus Grigorieff |
| author_facet | Johannes Elferich Marek Kaminek Lingli Kong Adolfo Odriozola Wanda Kukulski Benoît Zuber Nikolaus Grigorieff |
| author_sort | Johannes Elferich |
| collection | DOAJ |
| 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. |
| format | Article |
| id | doaj-art-c1f46eec874949e5a1b1b739865adf8b |
| institution | Matheson Library |
| issn | 2052-2525 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | International Union of Crystallography |
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| series | IUCrJ |
| spelling | doaj-art-c1f46eec874949e5a1b1b739865adf8b2025-07-03T10:13:26ZengInternational Union of CrystallographyIUCrJ2052-25252025-07-0112450251010.1107/S2052252525005196eh5022In situ high-resolution cryo-EM reconstructions from CEMOVISJohannes Elferich0Marek Kaminek1Lingli Kong2Adolfo Odriozola3Wanda Kukulski4Benoît Zuber5Nikolaus Grigorieff6Howard Hughes Medical Institute, University of Massachusetts Chan Medical School, Worcester, MA, USAInstitute of Anatomy, University of Bern, 3012 Bern, SwitzerlandRNA Therapeutics Institute, University of Massachusetts Chan Medical School, Worcester, MA, USAInstitute of Anatomy, University of Bern, 3012 Bern, SwitzerlandInstitute of Biochemistry and Molecular Medicine, University of Bern, 3012 Bern, SwitzerlandInstitute of Anatomy, University of Bern, 3012 Bern, SwitzerlandHoward Hughes Medical Institute, University of Massachusetts Chan Medical School, Worcester, MA, USACryo-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.https://journals.iucr.org/paper?S2052252525005196electron tomographyintegrative structural biologyimagingstructure determinationcryo-electron microscopy |
| spellingShingle | Johannes Elferich Marek Kaminek Lingli Kong Adolfo Odriozola Wanda Kukulski Benoît Zuber Nikolaus Grigorieff In situ high-resolution cryo-EM reconstructions from CEMOVIS IUCrJ electron tomography integrative structural biology imaging structure determination cryo-electron microscopy |
| title | In situ high-resolution cryo-EM reconstructions from CEMOVIS |
| title_full | In situ high-resolution cryo-EM reconstructions from CEMOVIS |
| title_fullStr | In situ high-resolution cryo-EM reconstructions from CEMOVIS |
| title_full_unstemmed | In situ high-resolution cryo-EM reconstructions from CEMOVIS |
| title_short | In situ high-resolution cryo-EM reconstructions from CEMOVIS |
| title_sort | in situ high resolution cryo em reconstructions from cemovis |
| topic | electron tomography integrative structural biology imaging structure determination cryo-electron microscopy |
| url | https://journals.iucr.org/paper?S2052252525005196 |
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