ArticleUltramicroscopy2014
Imaging endosomes and autophagosomes in whole mammalian cells using correlative cryo-fluorescence and cryo-soft X-ray microscopy (cryo-CLXM).
Article in Ultramicroscopy, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 55 papers, 2 of them syntheses that pooled it.
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Who cites it
55 citing papers in PubMed, 2 syntheses or guidelines pooled it.
- Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)Autophagy · 2021Guideline
- Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition).Autophagy · 2016Guideline
- Demonstrating soft X-ray tomography in the lab for correlative cryogenic biological imaging using X-rays and light microscopy.Scientific reports · 2025Article
- Cryo-CLXEM introduces cryo-SXT to bridge the resolution gap in cryo-CLEM.Communications biology · 2025Article
- Minimizing ice contamination during specimen preparation for cryo-soft X-ray tomography and cryo-electron tomography.Journal of structural biology: X · 2024Article
- Automated quantification of vacuole fusion and lipophagy inAutophagy · 2024Article
- Membrane association of the ATG8 conjugation machinery emerges as a key regulatory feature for autophagosome biogenesis.FEBS letters · 2024Review
- Kinetic modelling of sterol transport between plasma membrane and endo-lysosomes based on quantitative fluorescence and X-ray imaging data.Frontiers in cell and developmental biology · 2023Article
- Correlative Cryo-imaging Using Soft X-Ray Tomography for the Study of Virus Biology in Cells and Tissues.Sub-cellular biochemistry · 2023Article
- SHISA5/SCOTIN restrains spontaneous autophagy induction by blocking contact between the ERES and phagophores.Autophagy · 2022Article
- Microscopic Visualization of Cell-Cell Adhesion Complexes at Micro and Nanoscale.Frontiers in cell and developmental biology · 2022Review
- Single-Cell Chemistry of Photoactivatable Platinum Anticancer Complexes.Journal of the American Chemical Society · 2021Article
- Correlative 3D cryo X-ray imaging reveals intracellular location and effect of designed antifibrotic protein-nanomaterial hybrids.Chemical science · 2021Article
- Using soft X-ray tomography for rapid whole-cell quantitative imaging of SARS-CoV-2-infected cells.Cell reports methods · 2021Article
- Adhesion of Triple-Negative Breast Cancer Cells under Fluorescent and Soft X-ray Contact Microscopy.International journal of molecular sciences · 2021Article
- Sample preparation strategies for efficient correlation of 3D SIM and soft X-ray tomography data at cryogenic temperatures.Nature protocols · 2021Article
- Data-deposition protocols for correlative soft X-ray tomography and super-resolution structured illumination microscopy applications.STAR protocols · 2021Article
- Assaying three-dimensional cellular architecture using X-ray tomographic and correlated imaging approaches.The Journal of biological chemistry · 2020Review
- Mechanisms governing autophagosome biogenesis.Nature reviews. Molecular cell biology · 2020Review
- Article
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Authors and funding
9 authors.
Funding
Abstract
Cryo-soft X-ray tomography (cryo-SXT) is a powerful imaging technique that can extract ultrastructural information from whole, unstained mammalian cells as close to the living state as possible. Subcellular organelles including the nucleus, the Golgi apparatus and mitochondria have been identified by morphology alone, due to the similarity in contrast to transmission electron micrographs. In this study, we used cryo-SXT to image endosomes and autophagosomes, organelles that are particularly susceptible to chemical fixation artefacts during sample preparation for electron microscopy. We used two approaches to identify these compartments. For early and recycling endosomes, which are accessible to externally-loaded markers, we used an anti-transferrin receptor antibody conjugated to 10nm gold particles. For autophagosomes, which are not accessible to externally-applied markers, we developed a correlative cryo-fluorescence and cryo-SXT workflow (cryo-CLXM) to localise GFP-LC3 and RFP-Atg9. We used a stand-alone cryo-fluorescence stage in the home laboratory to localise the cloned fluorophores, followed by cryo-soft X-ray tomography at the synchrotron to analyse cellular ultrastructure. We mapped the 3D ultrastructure of the endocytic and autophagic structures, and discovered clusters of omegasomes arising from 'hotspots' on the ER. Thus, immunogold markers and cryo-CLXM can be used to analyse cellular processes that are inaccessible using other imaging modalities.
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Registered trials
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