ArticleScientific reports2019
Correlative cryo super-resolution light and electron microscopy on mammalian cells using fluorescent proteins.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 72 papers.
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Who cites it
72 citing papers in PubMed, 148 citations in OpenAlex.
- Challenges and limitations for live cell imaging in extreme cold.Methods and applications in fluorescence · 2026Article
- Cryoelectron tomography of HIV-1 cell-cell transmission conjugates reveals a secluded environment for viral assembly and transfer.bioRxiv : the preprint server for biology · 2026Article
- Integrated fluorescence light microscopy-guided cryo-focused ion beam-milling for in situ montage cryo-ET.Nature protocols · 2026Review
- A cryo-CLEM-guided workflow for quantitative assessment and optimization of cryo-ET specimen quality.Molecules and cells · 2026Article
- Fully genetically encoded low-molecular-weight protein tags with defined shapes for direct molecular identification by cryo-electron tomography.bioRxiv : the preprint server for biology · 2026Article
- Determination of factors that allow cryogenic nanoscopy with high power illumination without devitrification.PloS one · 2026Article
- Cryogenic light microscopy of vitrified samples with angstrom precision.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Mechanisms of COPII coat assembly and cargo recognition in the secretory pathway.Nature reviews. Molecular cell biology · 2025Review
- Single-Molecule 3D Nanoscopy for Application to Cryofixed Samples.The journal of physical chemistry. B · 2025Article
- Immunoelectron microscopy: a comprehensive guide from sample preparation to high-resolution imaging.Discover nano · 2025Review
- Cryo-light microscopy with angstrom precision deciphers structural conformations of PIEZO1 in its native state.Science advances · 2025Article
- Dissection of Neurochemical Pathways Across Complexity and Scale.Journal of neurochemistry · 2025Review
- A practical protocol for correlative confocal fluorescence and transmission electron microscopy characterization of extracellular vesicles.Microbiology spectrum · 2025Article
- IFITM2 Modulates Endocytosis Maintaining Neural Stem Cells in Developing Neocortex.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Unraveling atomic complexity from frozen samples.Structural dynamics (Melville, N.Y.) · 2025Article
- Cryogenic single-molecule fluorescence imaging.BMB reports · 2025Review
- AI-based hardware and software tools in microscopy to boost research in immunology and virology.Frontiers in immunology · 2025Review
- Understanding ubiquitination in neurodevelopment by integrating insights across space and time.Nature structural & molecular biology · 2025Review
- Recent advances in correlative cryo-light and electron microscopy.Current opinion in structural biology · 2024Review
- Exploring Transient States of PAmKate to Enable Improved Cryogenic Single-Molecule Imaging.Journal of the American Chemical Society · 2024Article
12 more citing papers are in PubMed but not listed here.
Corrections and comments
- Erratum issued
Authors and funding
5 authors at 3 institutions in 2 countries.
Funding
Abstract
Sample fixation by vitrification is critical for the optimal structural preservation of biomolecules and subsequent high-resolution imaging by cryo-correlative light and electron microscopy (cryoCLEM). There is a large resolution gap between cryo fluorescence microscopy (cryoFLM), ~400-nm, and the sub-nanometre resolution achievable with cryo-electron microscopy (cryoEM), which hinders interpretation of cryoCLEM data. Here, we present a general approach to increase the resolution of cryoFLM using cryo-super-resolution (cryoSR) microscopy that is compatible with successive cryoEM investigation in the same region. We determined imaging parameters to avoid devitrification of the cryosamples without the necessity for cryoprotectants. Next, we examined the applicability of various fluorescent proteins (FPs) for single-molecule localisation cryoSR microscopy and found that all investigated FPs display reversible photoswitchable behaviour, and demonstrated cryoSR on lipid nanotubes labelled with rsEGFP2 and rsFastLime. Finally, we performed SR-cryoCLEM on mammalian cells expressing microtubule-associated protein-2 fused to rsEGFP2 and performed 3D cryo-electron tomography on the localised areas. The method we describe exclusively uses commercially available equipment to achieve a localisation precision of 30-nm. Furthermore, all investigated FPs displayed behaviour compatible with cryoSR microscopy, making this technique broadly available without requiring specialised equipment and will improve the applicability of this emerging technique for cellular and structural biology.
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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.