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Instruments / Technical Area	Publication Details	Published	Actions
B24-Cryo Soft X-ray Tomography	Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells Mohamed A. Koronfel , Ilias Kounatidis , Dennis M. Mwangangi , Nina Vyas , Chidinma Okolo , Archana Jadhav , Tom Fish , Phatcharin Chotchuang , Albert Schulte , Robert Robinson , Maria Harkiolaki Acta Crystallographica Section D Structural Biology 77, 1479 - 1485 DOI: 10.1107/S2059798321010329 Diamond Proposal Number(s): [23033, 23073] Open Access Show Abstract ▼ Abstract: Imaging of actin filaments is crucial due to the integral role that they play in many cellular functions such as intracellular transport, membrane remodelling and cell motility. Visualizing actin filaments has so far relied on fluorescence microscopy and electron microscopy/tomography. The former lacks the capacity to capture the overall local ultrastructure, while the latter requires rigorous sample preparation that can lead to potential artefacts, and only delivers relatively small volumes of imaging data at the thinnest areas of a cell. In this work, a correlative approach utilizing in situ super-resolution fluorescence imaging and cryo X-ray tomography was used to image bundles of actin filaments deep inside cells under near-native conditions. In this case, fluorescence 3D imaging localized the actin bundles within the intracellular space, while X-ray tomograms of the same areas provided detailed views of the local ultrastructure. Using this new approach, actin trails connecting vesicles in the perinuclear area and hotspots of actin presence within and around multivesicular bodies were observed. The characteristic prevalence of filamentous actin in cytoplasmic extensions was also documented.	Dec 2021
B24-Cryo Soft X-ray Tomography	Protocol for image registration of correlative soft X-ray tomography and super-resolution structured illumination microscopy images Nina Vyas , Stephan Kunne , Thomas M. Fish , Ian M. Dobbie , Maria Harkiolaki , Perrine Paul-Gilloteaux Star Protocols 2 DOI: 10.1016/j.xpro.2021.100529 Diamond Proposal Number(s): [21046, 18314] Open Access Show Abstract ▼ Abstract: Correlation of 3D images acquired on different microscopes can be a daunting prospect even for experienced users. This protocol describes steps for registration of images from soft X-ray absorption contrast imaging and super-resolution fluorescence imaging of hydrated biological materials at cryogenic temperatures. Although it is developed for data generated at synchrotron beamlines that offer the above combination of microscopies, it is applicable to all analogous imaging systems where the same area of a sample is examined using successive non-destructive imaging techniques.	Jun 2021

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