ArticleScientific reports2023
CryoFIB milling large tissue samples for cryo-electron tomography.
Article in Scientific reports, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 18 citations in OpenAlex.
- Plant heteropolysaccharides and their bioadhesion to animal glycocalyx visualized at the nanoscale.Communications biology · 2026Article
- The Role of Confinement in Biomineralization.Chemical reviews · 2025Review
- Cryoprotectants-assisted plunge freezing of thick brain tissue specimens for targeted physiologically relevant cryo-imagingbioRxiv : the preprint server for biology · 2025Article
- TomoCPT: a generalizable model for 3D particle detection and localization in cryo-electron tomograms.Acta crystallographica. Section D, Structural biology · 2025Article
- Structural biology inside multicellular specimens using electron cryotomography.Quarterly reviews of biophysics · 2025Review
- Visualizing the virus world inside the cell by cryo-electron tomography.Journal of virology · 2024Review
- Missing Wedge Completion via Unsupervised Learning with Coordinate Networks.International journal of molecular sciences · 2024Article
- Missing Wedge Completion via Unsupervised Learning with Coordinate Networks.bioRxiv : the preprint server for biology · 2024Article
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Authors and funding
7 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cryo-electron tomography (cryoET) is a powerful tool for exploring the molecular structure of large organisms. However, technical challenges still limit cryoET applications on large samples. In particular, localization and cutting out objects of interest from a large tissue sample are still difficult steps. In this study, we report a sample thinning strategy and workflow for tissue samples based on cryo-focused ion beam (cryoFIB) milling. This workflow provides a full solution for isolating objects of interest by starting from a millimeter-sized tissue sample and ending with hundred-nanometer-thin lamellae. The workflow involves sample fixation, pre-sectioning, a two-step milling strategy, and localization of the object of interest using cellular secondary electron imaging (CSEI). The milling strategy consists of two steps, a coarse milling step to improve the milling efficiency, followed by a fine milling step. The two-step milling creates a furrow-ridge structure with an additional conductive Pt layer to reduce the beam-induced charging issue. CSEI is highlighted in the workflow, which provides on-the-fly localization during cryoFIB milling. Tests of the complete workflow were conducted to demonstrate the high efficiency and high feasibility of the proposed method.
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Registered trials
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