Abstract
Cryo-soft X-ray tomography is an imaging technique that addresses the need for mesoscale imaging of cellular ultrastructure of relatively thick samples without the need for staining or chemical modification. It allows the imaging of cellular ultrastructure to a resolution of 25-40 nm and can be used in correlation with other imaging modalities, such as electron tomography and fluorescence microscopy, to further enhance the information content derived from biological samples. An overview of the technique, discussion of sample suitability and information about sample preparation, data collection and data analysis is presented here. Recent developments and future outlook are also discussed.
🔬 Techniques
🧬 Organisms
🧪 Sample Preparation
🔬 Cell Lines
🏭 Microscope Brands
📷 Detectors
💻 Software Details
🏛️ Research Organizations (ROR)
Affiliated research institutions:
📊 Figures
Figureu00a01.
Biological imaging in context.
Representative techniques developed for the investigation of structure in biological systems at different scales, in order of resolution attainable.
Figureu00a02.
An example of typical cryo-SXT data.
( A ) A cryo-STX projection image of a Toxoplasma gondii containing vacuole within a human fibroblast cell. The reconstructed volume ( B ) shows the parasitophorous vacuole in yellow containing four p...
Figureu00a03.
A schematic of beamline B24 at the Diamond Light Source synchrotron.
X-rays are produced at a bending magnet, focused by a toroidal mirror and conditioned with a plane grating monochromator resulting in a secondary source at the exit slits. X-rays from his source are f...
Figureu00a04.
Cryo-SXT workflow.
Figureu00a05.
Typical cryo-STX imaging progression for adherent cells with fluorescence labels.
( A ) Brightfield grid overview of cryo-preserved mouse primary neuronal cells on a TEM grid (baru2009=u2009500u2005u00b5m), ( B ) close-up fluorescence and ( C ) brightfield images of the area on A m...
Figureu00a06.
Typical cryo-STX imaging workflow for smaller samples ( Trypanosome brucei ) with no associated fluorescence.
( A ) Brightfield overview of cryo-preserved cells on an area of holey carbon on a TEM grid (baru2009=u200950u2005u00b5m), ( B ) X-ray mosaic of the area on A marked by a green square (baru2009=u20092...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
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