Using the Volta phase plate with defocus for cryo-EM single particle analysis

被引：88

作者：

Danev, Radostin ^{[1
]}

Tegunov, Dimitry ^{[2
]}

Baumeister, Wolfgang ^{[1
]}

机构：

[1] Max Planck Inst Biochem, Dept Mol Struct Biol, Martinsried, Germany

[2] Max Planck Inst Biophys Chem, Dept Mol Biol, Gottingen, Germany

来源：

ELIFE | 2017年 / 6卷

关键词：

ELECTRON; ICE;

D O I：

10.7554/eLife.23006

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Previously, we reported an in-focus data acquisition method for cryo-EM single-particle analysis with the Volta phase plate (Danev and Baumeister, 2016). Here, we extend the technique to include a small amount of defocus which enables contrast transfer function measurement and correction. This hybrid approach simplifies the experiment and increases the data acquisition speed. It also removes the resolution limit inherent to the in-focus method thus allowing 3D reconstructions with resolutions better than 3 angstrom.

引用

页数：9

共 19 条

[1] 2.8 Å resolution reconstruction of the Thermoplasma acidophilum 20S proteasome using cryo-electron microscopy [J].

Campbell, Melody G. ;

Veesler, David ;

Cheng, Anchi ;

Potter, Clinton S. ;

Carragher, Bridget .

ELIFE, 2015, 4 :1-22

[2] Movies of Ice-Embedded Particles Enhance Resolution in Electron Cryo-Microscopy [J].

Campbell, Melody G. ;

Cheng, Anchi ;

Brilot, Axel F. ;

Moeller, Arne ;

Lyumkis, Dmitry ;

Veesler, David ;

Pan, Junhua ;

Harrison, Stephen C. ;

Potter, Clinton S. ;

Carragher, Bridget ;

Grigorieff, Nikolaus .

STRUCTURE, 2012, 20 (11) :1823-1828

[3] A Primer to Single-Particle Cryo-Electron Microscopy [J].

Cheng, Yifan ;

Grigorieff, Nikolaus ;

Penczek, Pawel A. ;

Walz, Thomas .

CELL, 2015, 161 (03) :438-449

[4] 3.9 Å structure of the nucleosome core particle determined by phase-plate cryo-EM [J].

Chua, Eugene Y. D. ;

Vogirala, Vinod K. ;

Inian, Oviya ;

Wong, Andrew S. W. ;

Nordenskiold, Lars ;

Plitzko, Juergen M. ;

Danev, Radostin ;

Sandin, Sara .

NUCLEIC ACIDS RESEARCH, 2016, 44 (17) :8013-8019

[5] Cryo-EM single particle analysis with the Volta phase plate [J].

Danev, Radostin ;

Baumeister, Wolfgang .

ELIFE, 2016, 5

[6] Volta potential phase plate for in-focus phase contrast transmission electron microscopy [J].

Danev, Radostin ;

Buijsse, Bart ;

Khoshouei, Maryam ;

Plitzko, Juergen M. ;

Baumeister, Wolfgang .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2014, 111 (44) :15635-15640

[7] Automatic estimation and correction of anisotropic magnification distortion in electron microscopes [J].

Grant, Timothy ;

Grigorieff, Nikolaus .

JOURNAL OF STRUCTURAL BIOLOGY, 2015, 192 (02) :204-208

[8] Bsoft: Image processing and molecular modeling for electron microscopy [J].

Heymann, J. Bernard ;

Belnap, David M. .

JOURNAL OF STRUCTURAL BIOLOGY, 2007, 157 (01) :3-18

[9]

Khoshouei M., 2016, BIORXIV, DOI [10.1101/087841, DOI 10.1101/087841]

[10] Volta phase plate cryo-EM of the small protein complex Prx3 [J].

Khoshouei, Maryam ;

Radjainia, Mazdak ;

Phillips, Amy J. ;

Gerrard, Juliet A. ;

Mitra, Alok K. ;

Plitzko, Juergen M. ;

Baumeister, Wolfgang ;

Danev, Radostin .

NATURE COMMUNICATIONS, 2016, 7

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