title: Cellular electron cryo tomography and in situ sub-volume averaging reveal the context of microtubule-based processes.
abstract: Electron cryo-tomography (cryoET) is currently the only technique that allows the direct observation of proteins in their native cellular environment. Sub-volume averaging of electron tomograms offers a route to increase the signal-to-noise of repetitive biological structures, such improving the information content and interpretability of tomograms. We discuss the potential for sub-volume averaging in highlighting and investigating specific processes in situ, focusing on microtubule structure and viral infection. We show that (i) in situ sub-volume averaging from single tomograms can guide and complement segmentation of biological features, (ii) the in situ determination of the structure of individual viruses is possible as they infect a cell, and (iii) novel, transient processes can be imaged with high levels of detail.
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, OX3 7BN, United Kingdom.
authors.forename: Michael
authors.initials: M
authors.lastname: Grange
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, OX3 7BN, United Kingdom.
authors.forename: Daven
authors.initials: D
authors.lastname: Vasishtan
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, OX3 7BN, United Kingdom. Electronic address: kay@strubi.ox.ac.uk.
authors.forename: Kay
authors.initials: K
authors.lastname: Grünewald
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D003599:Cytoskeleton / Q000648:ultrastructure; D004398:Dyneins / Q000648:ultrastructure; D055032:Electron Microscope Tomography / Q000379:methods*; D004705:Endocytosis / Q000502:physiology; D008870:Microtubules / Q000648:ultrastructure*
chemical_list: D004398:Dyneins
keywords: Adenovirus; Cellular architecture; Cytoskeleton; Dynein; Electron cryo-microscopy; Electron cryo-tomography; Endocytosis; Microtubules; Retrograde transport; Sub-volume averaging; Viral entry; Virus trafficking; in situ structure determination