Cryo Soft X-ray Tomography
Cryo Soft X-ray Tomography - Related Videos
08:47
A 3D Cartographic Description of the Cell by Cryo Soft X-ray Tomography
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Cited by 18 •
2021
Here, a protocol describing the sample preparation and data collection steps required in cryo soft X-ray tomography (SXT) to image the ultrastructure of whole cryo-preserved cells at a resolution of 25 nm half pitch, is presented.
06:56
Tree Core Analysis with X-ray Computed Tomography
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Cited by 10 •
2023
Here we show how to process tree cores with an X-ray computed tomography toolchain. Except for chemical extraction for some purposes, no further physical lab treatment is needed. The toolchain can be used for biomass estimations, for obtaining MXD/tree-ring width data as well as for obtaining quantitative wood anatomy data.
10:39
Visualization of ATP Synthase Dimers in Mitochondria by Electron Cryo-tomography
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Cited by 43 •
2014
We present a protocol of how to collect and process electron cryo-tomograms of whole mitochondria. The technique provides detailed insights into the structure, function, and organization of large membrane protein complexes in native biological membranes.
11:33
Using Tomoauto: A Protocol for High-throughput Automated Cryo-electron Tomography
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Cited by 48 •
2016
We present a protocol on how to utilize high-throughput cryo-electron tomography to determine high resolution in situ structures of molecular machines. The protocol permits large amounts of data to be processed, avoids common bottlenecks and reduces resource downtime, allowing the user to focus on important biological questions.
10:00
Energy Dispersive X-ray Tomography for 3D Elemental Mapping of Individual Nanoparticles
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Cited by 11 •
2016
The use of energy dispersive X-ray tomography in the scanning transmission electron microscope to characterize elemental distributions within single nanoparticles in three dimensions is described.
FAQs
Different cellular structures absorb soft X-rays to different extents, producing intensity differences in the transmitted projections. Computational reconstruction converts these absorption patterns into spatially organized features, allowing membranes, organelles, and inclusions to be distinguished without adding stains. This intrinsic contrast is particularly useful for examining cellular organization in frozen-hydrated specimens.
A single projection records only the structures encountered along one beam direction. Rotating the vitrified specimen collects projections from multiple angles, while computational reconstruction combines those measurements into a three-dimensional volume. The resulting dataset reveals how cellular components are arranged relative to one another, rather than providing only a two-dimensional view.
The method preserves intact, frozen-hydrated cells and uses their natural X-ray absorption contrast, so sectioning and staining are not required. Avoiding these steps helps retain broad cellular architecture while providing nanometer-scale structural detail. Researchers can therefore examine organelles, membranes, and inclusions within their surrounding cellular context instead of as isolated or processed structures.
Vitrification freezes the specimen in a frozen-hydrated state that supports imaging of cellular structures in a near-native condition. This preparation is important because the technique is intended to preserve intact cells rather than rely on sectioned or stained material. The preserved state enables structural observations to be interpreted alongside the overall organization of the cell.
A reconstructed volume can show the three-dimensional distribution of organelles, membranes, and inclusions across an intact cell. Because these features remain embedded within the broader cellular architecture, researchers can relate individual structures to their spatial surroundings. This combination of structural detail and whole-cell context supports analysis of cellular organization and changes associated with biological processes.
The technique is useful when researchers need structural information from intact cells while maintaining broad cellular context. In biology, applications include investigating cell organization, infection, development, and disease mechanisms. It can also help examine pathogen-host interactions by showing pathogens or associated cellular changes in relation to the surrounding host-cell architecture.