ArticleMicroscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada2019
Fluorescence-Based Detection of Membrane Fusion State on a Cryo-EM Grid using Correlated Cryo-Fluorescence and Cryo-Electron Microscopy.
Article in Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Integrated fluorescence light microscopy-guided cryo-focused ion beam-milling for in situ montage cryo-ET.Nature protocols · 2026Review
- Automated Cryo-EM and Supervised Machine Learning Enable Reproducible Characterization of Extracellular Vesicles and Co-Isolating Particles.Journal of extracellular vesicles · 2026Article
- Review
- Time-Resolved Fluorescence Imaging and Correlative Cryo-Electron Tomography to Study Structural Changes of the HIV-1 Capsid.ACS nano · 2025Article
- Recent advances in correlative cryo-light and electron microscopy.Current opinion in structural biology · 2024Review
- The Art of Viral Membrane Fusion and Penetration.Sub-cellular biochemistry · 2023Article
- Metallic support films reduce optical heating in cryogenic correlative light and electron tomography.Journal of structural biology · 2022Article
- Identification and demonstration of roGFP2 as an environmental sensor for cryogenic correlative light and electron microscopy.Journal of structural biology · 2022Article
- Sample preparation strategies for efficient correlation of 3D SIM and soft X-ray tomography data at cryogenic temperatures.Nature protocols · 2021Article
- Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies.Communications biology · 2021Article
- In Situ Imaging and Structure Determination of Biomolecular Complexes Using Electron Cryo-Tomography.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Human parainfluenza virus fusion complex glycoproteins imaged in action on authentic viral surfaces.PLoS pathogens · 2020Article
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Authors and funding
2 authors.
Funding
Abstract
Correlated light and electron microscopy (CLEM) has become a popular technique for combining the protein-specific labeling of fluorescence with electron microscopy, both at room and cryogenic temperatures. Fluorescence applications at cryo-temperatures have typically been limited to localization of tagged protein oligomers due to known issues of extended triplet state duration, spectral shifts, and reduced photon capture through cryo-CLEM objectives. Here, we consider fluorophore characteristics and behaviors that could enable more extended applications. We describe how dialkylcarbocanine DiD, and its autoquenching by resonant energy transfer (RET), can be used to distinguish the fusion state of a lipid bilayer at cryo-temperatures. By adapting an established fusion assay to work under cryo-CLEM conditions, we identified areas of fusion between influenza virus-like particles and fluorescently labeled lipid vesicles on a cryo-EM grid. This result demonstrates that cryo-CLEM can be used to localize functions in addition to tagged proteins, and that fluorescence autoquenching by RET can be incorporated successfully into cryo-CLEM approaches. In the case of membrane fusion applications, this method provides both an orthogonal confirmation of functional state independent of the morphological description from cryo-EM and a way to bridge room-temperature kinetic assays and the cryo-EM images.
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