2025/06/19 by Johannes Elferich, Marek Kaminek, Lingli Kong +4 · 1 voice
Biochemistry, Genetics and Molecular Biology · Engineering · Materials Science · #Advanced Electron Microscopy Techniques and Applications #Electron and X-Ray Spectroscopy Techniques #Integrated Circuits and Semiconductor Failure Analysis
paper · pdf · doi:10.1107/s2052252525005196
openalex publication_date 2025/06/19 · openalex created_date 2025/06/24 · openalex updated_date 2026/07/31
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.