ArticleMethods in molecular biology (Clifton, N.J.)2021
Practical Approaches for Cryo-FIB Milling and Applications for Cellular Cryo-Electron Tomography.
Article in Methods in molecular biology (Clifton, N.J.), 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed.
- Article
- Applications and prospects of cryo-electron tomography in drug discovery and understanding disease.Current opinion in structural biology · 2026Review
- Integrated fluorescence light microscopy-guided cryo-focused ion beam-milling for in situ montage cryo-ET.Nature protocols · 2026Review
- The phage nucleus synergizes with an anti-defense protein to resist bacterial immunity.Cell reports · 2026Article
- Molecular architecture of the ciliary base in mammalian multiciliated cells.bioRxiv : the preprint server for biology · 2026Article
- A cryo-CLEM-guided workflow for quantitative assessment and optimization of cryo-ET specimen quality.Molecules and cells · 2026Article
- Human NLRP3 inflammasome activation leads to formation of condensate at the microtubule organizing center.Science advances · 2026Article
- ExoSloNano: multimodal nanogold labels for identification of macromolecules in live cells and cryo-electron tomograms.Nature methods · 2026Article
- Article
- Workflow for Fluorescence-Targeted Lamella Milling From Vitrified Cells With a Coincident Fluorescence, Electron, and Ion Beam Microscope.Bio-protocol · 2025Article
- Sequential membrane- and protein-bound organelles compartmentalize genomes during phage infection.Cell host & microbe · 2025Article
- Mapping the ultrastructural topology of the corynebacterial cell surface.PLoS biology · 2025Article
- A new statistical metric for robust target detection in cryo-EM using 2D template matching.IUCrJ · 2025Article
- Rapid structural analysis of bacterial ribosomes in situ.Communications biology · 2025Article
- Leveraging Virtual Reality for Immersive Segmentation and Analysis of Cryo-Electron Tomography Data.Journal of visualized experiments : JoVE · 2025Article
- Unravelling complex mechanisms in materials processes with cryogenic electron microscopy.Chemical science · 2025Review
- Structural biology inside multicellular specimens using electron cryotomography.Quarterly reviews of biophysics · 2025Review
- Spatial Proteomics towards cellular Resolution.Expert review of proteomics · 2024Article
- A transcriptionally active lipid vesicle encloses the injectedbioRxiv : the preprint server for biology · 2024Article
- Article
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Abstract
Cryo-electron tomography (cryo-ET) is a powerful technique to examine cellular structures as they exist in situ. However, direct imaging by TEM for cryo-ET is limited to specimens up to ∼400 nm in thickness, narrowing its applicability to areas such as cellular projections or small bacteria and viruses. Cryo-focused ion beam (cryo-FIB) milling has emerged in recent years as a method to generate thin specimens from cellular samples in preparation for cryo-ET. In this technique, specimens are thinned with a beam of gallium ions to gradually ablate cellular material in order to leave a thin, electron-transparent section (a lamella) through the bulk material. The lamella can be used for high-resolution cryo-ET to visualize cells in 3D in a near-native state. This approach has proved to be robust and relatively simple for new users and exhibits minimal sectioning artifacts. In this chapter, we describe a general approach to cryo-FIB milling for users with prior cryo-EM experience, with extensive notes on operation and troubleshooting.
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