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.
J Struct Biol
02/2017
197
181 - 190
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, Cryoelectron Microscopy, Cytoskeleton, Dyneins, Electron Microscope Tomography, Endocytosis, Microtubules