Please use this identifier to cite or link to this item: https://doi.org/10.7554/eLife.34257
Title: Routine single particle cryoEM sample and grid characterization by tomography
Authors: Noble, Alex J
Dandey, Venkata P
Wei, Hui
Braschi, Julia
Chase, Jillian
Acharya, Priyamvada
Tan, Yong Zi 
Zhang, Zhening
Kim, Laura Y
Scapin, Giovanna
Rapp, Micah
Eng, Edward T
Rice, William J
Cheng, Anchi
Negro, Carl J
Shapiro, Lawrence
Kwong, Peter D
Jeruzalmi, David
des Georges, Amedee
Potter, Clinton S
Carragher, Bridget
Keywords: Science & Technology
Life Sciences & Biomedicine
Biology
Life Sciences & Biomedicine - Other Topics
AIR-WATER-INTERFACE
EM STRUCTURE DETERMINATION
ATOMIC-FORCE MICROSCOPY
BEAM-INDUCED MOTION
HOLEY CARBON-FILM
CRYOELECTRON MICROSCOPY
AIR/WATER INTERFACE
ELECTRON-MICROSCOPY
PROTEIN ADSORPTION
HIGH-RESOLUTION
Issue Date: 29-May-2018
Publisher: ELIFE SCIENCES PUBLICATIONS LTD
Citation: Noble, Alex J, Dandey, Venkata P, Wei, Hui, Braschi, Julia, Chase, Jillian, Acharya, Priyamvada, Tan, Yong Zi, Zhang, Zhening, Kim, Laura Y, Scapin, Giovanna, Rapp, Micah, Eng, Edward T, Rice, William J, Cheng, Anchi, Negro, Carl J, Shapiro, Lawrence, Kwong, Peter D, Jeruzalmi, David, des Georges, Amedee, Potter, Clinton S, Carragher, Bridget (2018-05-29). Routine single particle cryoEM sample and grid characterization by tomography. ELIFE 7. ScholarBank@NUS Repository. https://doi.org/10.7554/eLife.34257
Abstract: Single particle cryo-electron microscopy (cryoEM) is often performed under the assumption that particles are not adsorbed to the air-water interfaces and in thin, vitreous ice. In this study, we performed fiducial-less tomography on over 50 different cryoEM grid/sample preparations to determine the particle distribution within the ice and the overall geometry of the ice in grid holes. Surprisingly, by studying particles in holes in 3D from over 1000 tomograms, we have determined that the vast majority of particles (approximately 90%) are adsorbed to an air-water interface. The implications of this observation are wide-ranging, with potential ramifications regarding protein denaturation, conformational change, and preferred orientation. We also show that fiducial-less cryo-electron tomography on single particle grids may be used to determine ice thickness, optimal single particle collection areas and strategies, particle heterogeneity, and de novo models for template picking and single particle alignment.
Source Title: ELIFE
URI: https://scholarbank.nus.edu.sg/handle/10635/227212
ISSN: 2050084X
DOI: 10.7554/eLife.34257
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