A Fast and Effective Microfluidic Spraying-Plunging Method for High-Resolution Single-Particle Cryo-EM.
Xiangsong Feng,Ziao Fu,Sandip Kaledhonkar,Yuan Jia,Binita Shah,Amy Jin,Zheng Liu,Ming Sun,Bo Chen,Robert A. Grassucci,Yukun Ren,Hongyuan Jiang,Joachim Frank,Joachim Frank,Qiao Lin +14 more
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TLDR
A spraying-plunging method for preparing cryoelectron microscopy grids with vitreous ice of controllable, highly consistent thickness using a microfluidic device is described and it is demonstrated that the structure can be solved to high resolution with this method of sample preparation.About:
This article is published in Structure.The article was published on 2017-04-04 and is currently open access. It has received 99 citations till now.read more
Citations
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Cryo-EM in drug discovery: achievements, limitations and prospects
Jean-Paul Renaud,Ashwin Chari,Claudio Ciferri,Wen-Ti Liu,Hervé-William Rémigy,Holger Stark,Christian Wiesmann +6 more
TL;DR: The recent advances in the field are described and critically assesses their relevance for drug discovery as well as discussing at what stages of the drug discovery pipeline cryo-EM can be useful today and what to expect in the near future.
Journal ArticleDOI
Challenges and opportunities in cryo-EM single-particle analysis
TL;DR: Some of the main methods surrounding cryo-EM with an emphasis specifically on single-particle analysis are reviewed, and challenges, open questions, and opportunities for methodology development are highlighted.
Journal ArticleDOI
Routine single particle CryoEM sample and grid characterization by tomography.
Alex J. Noble,Venkata P. Dandey,Hui Wei,Julia Brasch,Jillian Chase,Jillian Chase,Priyamvada Acharya,Yong Zi Tan,Zhening Zhang,Laura Y. Kim,Giovanna Scapin,Micah Rapp,Edward T. Eng,William J. Rice,Anchi Cheng,Carl J. Negro,Lawrence Shapiro,Peter D. Kwong,David Jeruzalmi,Amedee des Georges,Clinton S. Potter,Bridget Carragher +21 more
TL;DR: Surprisingly, by studying particles in holes in 3D from over 1000 tomograms, it is determined that the vast majority of particles (approximately 90%) are adsorbed to an air-water interface.
Journal ArticleDOI
Reducing effects of particle adsorption to the air–water interface in cryo-EM
Alex J. Noble,Hui Wei,Venkata P. Dandey,Zhening Zhang,Yong Zi Tan,Clinton S. Potter,Bridget Carragher +6 more
TL;DR: Reducing the length of time that protein particles spend on a sample grid prior to freezing mitigates deleterious effects caused by particle adsorption to the air–water interface in single-particle cryo-EM.
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Spotiton: New features and applications.
Venkata P. Dandey,Hui Wei,Zhening Zhang,Yong Zi Tan,Priyamvada Acharya,Edward T. Eng,William J. Rice,Peter A. Kahn,Clinton S. Potter,Bridget Carragher +9 more
TL;DR: Spotiton has been used to prepare several test specimens that can be reconstructed using routine single particle analysis to ∼3 Å resolution, indicating that the process has no apparent deleterious effect on the sample integrity.
References
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Features and development of Coot.
TL;DR: Coot is a molecular-graphics program designed to assist in the building of protein and other macromolecular models and the current state of development and available features are presented.
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RELION: implementation of a Bayesian approach to cryo-EM structure determination.
TL;DR: Developments that reduce the computational costs of the underlying maximum a posteriori (MAP) algorithm, as well as statistical considerations that yield new insights into the accuracy with which the relative orientations of individual particles may be determined are described.
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CTFFIND4: Fast and accurate defocus estimation from electron micrographs.
Alexis Rohou,Nikolaus Grigorieff +1 more
TL;DR: Modifications to the CTFFIND algorithm are described which make it significantly faster and more suitable for use with images collected using modern technologies such as dose fractionation and phase plates.
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Electron counting and beam-induced motion correction enable near-atomic-resolution single-particle cryo-EM
Xueming Li,Paul Mooney,Shawn Q. Zheng,Shawn Q. Zheng,Christopher R. Booth,Michael B. Braunfeld,Michael B. Braunfeld,Sander Gubbens,David A. Agard,David A. Agard,Yifan Cheng +10 more
TL;DR: This approach determined a 3.3-Å-resolution structure of an ∼700-kDa protein with D7 symmetry, the Thermoplasma acidophilum 20S proteasome, showing clear side-chain density and greatly enhances image quality and data acquisition efficiency.
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Cryo-electron microscopy of viruses
TL;DR: Cryo-electron microscopy of vitrified specimens offers possibilities for high resolution observations that compare favourably with any other electron microscopical method.