ArticleNature protocols2021
Sample preparation strategies for efficient correlation of 3D SIM and soft X-ray tomography data at cryogenic temperatures.
Article in Nature protocols, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed.
- Radiation-dose effects in correlative X-ray/cryo-electron microscopy of frozen-hydrated biological samples.Acta crystallographica. Section D, Structural biology · 2026Article
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- µCodes: A Universal Grid Platform for Microscale Mapping, Microscopy Navigation, and Multimodal Imaging.Small (Weinheim an der Bergstrasse, Germany) · 2025Article
- Advances in automation for cryo-electron tomography data collection.Current opinion in structural biology · 2025Review
- Cryo-CLXEM introduces cryo-SXT to bridge the resolution gap in cryo-CLEM.Communications biology · 2025Article
- Internalization and Cellular Fate of Protein Corona-Coated Nanoparticles by Multimodal Multi-Scale Microscopy.Small (Weinheim an der Bergstrasse, Germany) · 2025Article
- Sample Preparation Protocol for Laboratory Cryo-Soft X-Ray Microscopy for Studying Cellular Nanoparticle Uptake.International journal of molecular sciences · 2025Article
- Soft X-ray tomography analysis of mitochondria dynamics in Saccharomyces cerevisiae.Biology direct · 2024Article
- Correlative light and soft X-ray tomography of in situ mesoscale heterochromatin structure in intact cells.Scientific reports · 2024Article
- Article
- Automated quantification of vacuole fusion and lipophagy inAutophagy · 2024Article
- Article
- Laboratory Liquid-Jet X-ray Microscopy and X-ray Fluorescence Imaging for Biomedical Applications.International journal of molecular sciences · 2024Article
- In situ label-free X-ray imaging for visualizing the localization of nanomedicines and subcellular architecture in intact single cells.Nature protocols · 2024Review
- On-Line Fluorescence Microscopy for Identification and Imaging of Apoptotic Cell with Synchrotron-Based Soft X-ray Tomography.Micromachines · 2023Article
- Correlative Cryo-imaging Using Soft X-Ray Tomography for the Study of Virus Biology in Cells and Tissues.Sub-cellular biochemistry · 2023Article
- 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
- Near-native state imaging by cryo-soft-X-ray tomography reveals remodelling of multiple cellular organelles during HSV-1 infection.PLoS pathogens · 2022Article
- A guide into the world of high-resolution 3D imaging: the case of soft X-ray tomography for the life sciences.Biochemical Society transactions · 2022Article
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Authors and funding
11 authors.
Funding
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Abstract
3D correlative microscopy methods have revolutionized biomedical research, allowing the acquisition of multidimensional information to gain an in-depth understanding of biological systems. With the advent of relevant cryo-preservation methods, correlative imaging of cryogenically preserved samples has led to nanometer resolution imaging (2-50 nm) under harsh imaging regimes such as electron and soft X-ray tomography. These methods have now been combined with conventional and super-resolution fluorescence imaging at cryogenic temperatures to augment information content from a given sample, resulting in the immediate requirement for protocols that facilitate hassle-free, unambiguous cross-correlation between microscopes. We present here sample preparation strategies and a direct comparison of different working fiducialization regimes that facilitate 3D correlation of cryo-structured illumination microscopy and cryo-soft X-ray tomography. Our protocol has been tested at two synchrotron beamlines (B24 at Diamond Light Source in the UK and BL09 Mistral at ALBA in Spain) and has led to the development of a decision aid that facilitates experimental design with the strategic use of markers based on project requirements. This protocol takes between 1.5 h and 3.5 d to complete, depending on the cell populations used (adherent cells may require several days to grow on sample carriers).
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