3D images of biological cells at a resolution superior to the light microscope with specimens much thicker than possible with electron microscopy
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The main steps for x-ray imaging are specimen preparation, x-ray data collection, data processing and analysis of the images.
Specimen preparation is similar to that for cryo-electron microscopy but the samples are usually thicker. Care has to be taken to avoid freezing artefacts. We provide facilities within Instruct for specimen preparation at the x-ray imaging synchrotron beamlines and elsewhere. X-ray data collection takes place at the synchrotron beamlines and facilities for data processing and analysis are also available both at the beamlines and other Instruct centres.
Full field imaging microscopes are equipped with zone plates as objective lenses. At present these work within the "water window" at approximately 500eV where maximum absorption contrast is provided. Fields of view of approximately 20µm are typical with specimen thicknesses of 5µm. Operation in phase contrast at higher energies will allow thicker specimens to be examined. These microscopes are the most established for structural cell biology.
Scanning microscopes provide a small focal spot which scans across the specimen building up the two dimensional images. These operate at higher energies and are therefore suitable for use in the investigation of thicker biological cells and tissues. They include the option of capturing x-ray fluorescence in combination with differential phase contrast images thus enabling the positioning of heavier elements with respect to cellular organelles.