ArticleBiophysical journal2016
Site-Specific Cryo-focused Ion Beam Sample Preparation Guided by 3D Correlative Microscopy.
Article in Biophysical journal, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 96 papers.
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Who cites it
96 citing papers in PubMed, 231 citations in OpenAlex.
- Optimized workflow of site-specific cryo-lamella preparation for cryo-ET using integrated light and electron microscope.Biophysics reports · 2026Article
- Integrated fluorescence light microscopy-guided cryo-focused ion beam-milling for in situ montage cryo-ET.Nature protocols · 2026Review
- Electron microscopy-based three-dimensional subcellular imaging of plant male gametophyte.Journal of integrative plant biology · 2026Review
- Neurodegeneration risk variants promote lysosomal TMEM106B fibril accumulation.bioRxiv : the preprint server for biology · 2026Article
- Human NLRP3 inflammasome activation leads to formation of condensate at the microtubule organizing center.Science advances · 2026Article
- Optical interference for the guidance of cryogenic focused ion beam milling beyond the axial diffraction limit.Nature communications · 2026Article
- Article
- There and back again: a cell biologist's journey from organelles to molecules.Biological chemistry · 2025Review
- Cryo-CLXEM introduces cryo-SXT to bridge the resolution gap in cryo-CLEM.Communications biology · 2025Article
- Direct visualization of HIV-1 core nuclear import and its interplay with the nuclear pore.EMBO reports · 2025Article
- Cryoprotectants-assisted plunge freezing of thick brain tissue specimens for targeted physiologically relevant cryo-imagingbioRxiv : the preprint server for biology · 2025Article
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- Molecular Visualization of Neuronal TDP43 PathologybioRxiv : the preprint server for biology · 2025Article
- Morphology and mechanical behavior of diatoms in wet and dry states studied using nano-XCT.BMC biology · 2025Article
- In situ cryo-electron microscopy and tomography of cellular and organismal samples.Current opinion in structural biology · 2025Review
- HIV-1 nuclear import is selective and depends on both capsid elasticity and nuclear pore adaptability.Nature microbiology · 2025Article
- Workflow for Fluorescence-Targeted Lamella Milling From Vitrified Cells With a Coincident Fluorescence, Electron, and Ion Beam Microscope.Bio-protocol · 2025Article
- TomoScore: A Neural Network Approach for Quality Assessment of Cellular cryo-ET.Research square · 2025Article
- Structure of the lens MP20 mediated adhesive junction.Nature communications · 2025Article
- Thickness- and quality-controlled fabrication of fluorescence-targeted frozen-hydrated lamellae.Cell reports methods · 2025Article
36 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
10 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The development of cryo-focused ion beam (cryo-FIB) for the thinning of frozen-hydrated biological specimens enabled cryo-electron tomography (cryo-ET) analysis in unperturbed cells and tissues. However, the volume represented within a typical FIB lamella constitutes a small fraction of the biological specimen. Retaining low-abundance and dynamic subcellular structures or macromolecular assemblies within such limited volumes requires precise targeting of the FIB milling process. In this study, we present the development of a cryo-stage allowing for spinning-disk confocal light microscopy at cryogenic temperatures and describe the incorporation of the new hardware into existing workflows for cellular sample preparation by cryo-FIB. Introduction of fiducial markers and subsequent computation of three-dimensional coordinate transformations provide correlation between light microscopy and scanning electron microscopy/FIB. The correlative approach is employed to guide the FIB milling process of vitrified cellular samples and to capture specific structures, namely fluorescently labeled lipid droplets, in lamellas that are 300 nm thick. The correlation procedure is then applied to localize the fluorescently labeled structures in the transmission electron microscopy image of the lamella. This approach can be employed to navigate the acquisition of cryo-ET data within FIB-lamellas at specific locations, unambiguously identified by fluorescence microscopy.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.