Abstract
Imaging techniques in structural cell biology are indispensable to understand cell organization and machinery. In this frame, cryo soft X-ray tomography (cryo-SXT), a synchrotron-based imaging technique, is used to analyze the ultrastructure of intact, cryo-preserved cells at nanometric spatial resolution bridging electron microscopy and visible light fluorescence. With their unique interaction with matter and high penetration depth, X-rays are a very useful and complementary source to obtain both high-resolution and quantitative information. In this review, we are elaborating a typical cryo correlative workflow at the Mistral Beamline at the Alba Synchrotron (Spain) with the goal of providing a cartographic description of the cell by cryo-SXT that illustrates the possibilities this technique brings for specific localization of cellular features, organelle organization, and particular events in specific structural cell biology research.
Highlights
Structural biology has been one of the greatest beneficiaries from the development of advanced microscopic techniques
Like focused ion beam/ scanning electron microscopy (FIB-SEM) or serial block-face imaging, Biophys Rev (2019) 11:611–619 have greatly improved the automated acquisition of 3D information, making it much easier and faster
Cryo-capabilities have been developed so that cryo-FIB milling of the sample allows for cryo electron tomography (ET) avoiding staining and dehydration, but cryo-ET is still a non-routine technique compared with the classical transmission electron microscopy (TEM) on epoxy resin–embedded sections
Summary
Structural biology has been one of the greatest beneficiaries from the development of advanced microscopic techniques. Once the grids have been loaded into the TXM (4 samples at a time), a first grid is chosen and an overview of the different grid squares on which interesting positions have been previously localized by cryo-epifluorescence are imaged to find the specific cells These organelles are difficult to visualize because their membrane size is close to the resolution limit of the cryo-SXT (Mitra et al 2004). This is the easiest to recognize, it is only one of its possible forms
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