ArticleJournal of structural biology2017
New hardware and workflows for semi-automated correlative cryo-fluorescence and cryo-electron microscopy/tomography.
Article in Journal of structural biology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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Who cites it
36 citing papers in PubMed.
- Integrated fluorescence light microscopy-guided cryo-focused ion beam-milling for in situ montage cryo-ET.Nature protocols · 2026Review
- There and back again: a cell biologist's journey from organelles to molecules.Biological chemistry · 2025Review
- Time-Resolved Fluorescence Imaging and Correlative Cryo-Electron Tomography to Study Structural Changes of the HIV-1 Capsid.ACS nano · 2025Article
- Serial Lift-Out: sampling the molecular anatomy of whole organisms.Nature methods · 2024Article
- Correlative Imaging to Detect Rare HIV Reservoirs and Associated Damage in Tissues.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Recent advances and current trends in cryo-electron microscopy.Current opinion in structural biology · 2022Review
- Article
- Bimodal endocytic probe for three-dimensional correlative light and electron microscopy.Cell reports methods · 2022Article
- Sample preparation and image registration for correlative cryo-FM and cryo-FIB-SEM of plunge-frozen mammalian cells.STAR protocols · 2022Article
- Quantitative Cryo-Electron Tomography.Frontiers in molecular biosciences · 2022Review
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- Article
- CorRelator: Interactive software for real-time high precision cryo-correlative light and electron microscopy.Journal of structural biology · 2021Article
- HIV-1 uncoating by release of viral cDNA from capsid-like structures in the nucleus of infected cells.eLife · 2021Article
- Cryogenic Super-Resolution Fluorescence and Electron Microscopy Correlated at the Nanoscale.Annual review of physical chemistry · 2021Review
- Article
- Cryo-fluorescence microscopy of high-pressure frozen C. elegans enables correlative FIB-SEM imaging of targeted embryonic stages in the intact worm.Methods in cell biology · 2021Article
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- AutoCLEM: An Automated Workflow for Correlative Live-Cell Fluorescence Microscopy and Cryo-Electron Tomography.Scientific reports · 2019Article
- Advances in Cryo-Correlative Light and Electron Microscopy: Applications for Studying Molecular and Cellular Events.The protein journal · 2019Review
Corrections and comments
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Authors and funding
10 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Correlative light and electron microscopy allows features of interest defined by fluorescence signals to be located in an electron micrograph of the same sample. Rare dynamic events or specific objects can be identified, targeted and imaged by electron microscopy or tomography. To combine it with structural studies using cryo-electron microscopy or tomography, fluorescence microscopy must be performed while maintaining the specimen vitrified at liquid-nitrogen temperatures and in a dry environment during imaging and transfer. Here we present instrumentation, software and an experimental workflow that improves the ease of use, throughput and performance of correlated cryo-fluorescence and cryo-electron microscopy. The new cryo-stage incorporates a specially modified high-numerical aperture objective lens and provides a stable and clean imaging environment. It is combined with a transfer shuttle for contamination-free loading of the specimen. Optimized microscope control software allows automated acquisition of the entire specimen area by cryo-fluorescence microscopy. The software also facilitates direct transfer of the fluorescence image and associated coordinates to the cryo-electron microscope for subsequent fluorescence-guided automated imaging. Here we describe these technological developments and present a detailed workflow, which we applied for automated cryo-electron microscopy and tomography of various specimens.
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Registered trials
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