Abstract
One of the ultimate aims of imaging in biology is to achieve molecular localisation in the context of the structure of cells in their native state. Here, we review the current state of the art in cryo-soft X-ray tomography (cryo-SXT), which is the only imaging modality that can provide nanoscale 3D information from cryo-preserved, unstained, whole cells thicker than 1 μm. Correlative cryo-fluorescence and cryo-SXT adds functional information to structure, enabling studies of cellular events that cannot be captured using light, electron or X-ray microscopes alone.
Highlights
Recent years have seen an explosion of electron microscope technologies for high-resolution 3D imaging of biological samples at nanometre resolution (Fig. 1)
For samples embedded in resin, well-established transmission electron microscopy (TEM) techniques and, Handling Editor: David Robinson
This was especially important when moving from 2D to 3D imaging using cryo-Soft X-ray tomography (SXT) (Schneider 1998; Maser et al 2000; Wang et al 2000; Weiss et al 2000), where the sample is exposed to multiple doses during tilt series data collection
Summary
Recent years have seen an explosion of electron microscope technologies for high-resolution 3D imaging of biological samples at nanometre resolution (Fig. 1). The development of suitable optics such as grazing incidence mirrors, monochromators, and multilayer optics for energy selection and focusing of the intense X-ray beams, alongside developments in zone plate manufacturing (the optical component in the microscope that governs the overall achievable resolution, assuming a perfect detector and an ideal sample), increased the attraction of X-ray microscopy to instrument developers, technologists and research scientists interested in the ultimate application of the technique. Such was the momentum generated by these advances that a series of international conferences were organised focusing on the latest developments. Great care needs to be taken in setting up the microscope to ensure that there are no clashes with the sample mount as the sample is rotated during tomographic data collection
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