Removing high contrast artifacts via digital inpainting in cryo-electron tomography: An application of compressed sensing

2012 ◽  
Vol 178 (2) ◽  
pp. 108-120 ◽  
Author(s):  
Kahye Song ◽  
Luis R. Comolli ◽  
Mark Horowitz
Keyword(s):  
Compressed Sensing ◽  
Electron Tomography ◽  
High Contrast ◽  
Cryo Electron Tomography ◽  
Digital Inpainting

scholarly journals Dual-axis Volta phase plate cryo-electron tomography of Ebola virus-like particles reveals actin-VP40 interactions

2021 ◽  
Author(s):  
Sophie L. Winter ◽  
Petr Chlanda
Keyword(s):  
Actin Filaments ◽  
Ebola Virus ◽  
Electron Tomography ◽  
Direct Interaction ◽  
Viral Envelope ◽  
Phase Plate ◽  
High Contrast ◽  
Slab Geometry ◽  
Virus Like Particles ◽  
Cryo Electron Tomography

AbstractCryo-electron tomography (cryo-ET) is a pivotal imaging technique for studying the structure of pleomorphic enveloped viruses and their interactions with the host at native conditions. Owing to the limited tilting range of samples with a slab geometry, electron tomograms suffer from so-called missing wedge information in Fourier space. In dual-axis cryo-ET, two tomograms reconstructed from orthogonally oriented tilt series are combined into a tomogram with improved resolution as the missing wedge information is reduced to a pyramid. Volta phase plate (VPP) allows to perform in-focus cryo-ET with high contrast transfer at low-resolution frequencies and thus its application may improve the quality of dual-axis tomograms. Here, we compare dual-axis cryo-ET with and without VPP on Ebola virus-like particles to visualize and segment viral and host cell proteins within the membrane-enveloped filamentous particles. Dual-axis VPP cryo-ET reduces the missing wedge information and ray artifacts arising from the weighted back-projection during tomogram reconstruction, thereby minimizing ambiguity in the analysis of crowded environments and facilitating 3D segmentation. We show that dual-axis VPP tomograms provide a comprehensive description of macromolecular organizations such as nucleocapsid assembly states, the distribution of glycoproteins on the viral envelope and asymmetric arrangements of the VP40 layer in non-filamentous regions of virus-like particles. Our data reveal actin filaments within virus-like particles in close proximity to the viral VP40 scaffold, suggesting a direct interaction between VP40 and actin filaments. Dual-axis VPP cryo-ET provides more complete 3D information at high contrast and allows for better interpretation of macromolecule interactions and pleomorphic organizations.HighlightsVolta phase plate dual-axis cryo-electron tomography provides high contrast tomography data with reduced back-projection ray artifacts and missing wedge information in Fourier spaceVolta phase plate dual-axis cryo-electron tomography facilitates interpretation of protein-membrane interactionsVolta phase plate dual-axis cryo-electron tomography reduces ambiguity in manual 3D rendering and markedly improves 3D isosurface modelingEbola virus-like particles contain actin filaments in close proximity to the VP40 layer

scholarly journals Determining the Patchwork Lattice of Ebola and Marburg Virus Matrix Layers Using Cryo-Electron Tomography

2021 ◽  
Vol 27 (S1) ◽  
pp. 1884-1884
Author(s):  
William Wan ◽  
Mairi Clarke ◽  
Michael Norris ◽  
Larissa Kolesnikova ◽  
Alexander Koehler ◽  
...  
Keyword(s):  
Electron Tomography ◽  
Marburg Virus ◽  
Cryo Electron Tomography
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