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Author Tsai, C.-Y.; Chang, Y.-C.; Lobato, I.; Van Dyck, D.; Chen, F.-R.
Title Hollow Cone Electron Imaging for Single Particle 3D Reconstruction of Proteins Type A1 Journal article
Year (down) 2016 Publication Scientific reports Abbreviated Journal Sci Rep-Uk
Volume 6 Issue 6 Pages 27701
Keywords A1 Journal article; Electron microscopy for materials research (EMAT); Vision lab
Abstract The main bottlenecks for high-resolution biological imaging in electron microscopy are radiation sensitivity and low contrast. The phase contrast at low spatial frequencies can be enhanced by using a large defocus but this strongly reduces the resolution. Recently, phase plates have been developed to enhance the contrast at small defocus but electrical charging remains a problem. Single particle cryo-electron microscopy is mostly used to minimize the radiation damage and to enhance the resolution of the 3D reconstructions but it requires averaging images of a massive number of individual particles. Here we present a new route to achieve the same goals by hollow cone dark field imaging using thermal diffuse scattered electrons giving about a 4 times contrast increase as compared to bright field imaging. We demonstrate the 3D reconstruction of a stained GroEL particle can yield about 13.5 A resolution but using a strongly reduced number of images.
Address Department of Engineering and System Science, Tsing-Hua University, HsinChu 300, Taiwan
Corporate Author Thesis
Publisher Place of Publication Editor
Language English Wos 000377670500001 Publication Date 2016-06-13
Series Editor Series Title Abbreviated Series Title
Series Volume Series Issue Edition
ISSN 2045-2322 ISBN Additional Links UA library record; WoS full record; WoS citing articles
Impact Factor 4.259 Times cited Open Access
Notes D. Van Dyck acknowledges the financial support from the Fund for Scientific Research – Flanders (FWO) under Project nos. VF04812N and G.0188.08. F. R. Chen would like to thank the support from NSC 101-2221-E-007- 063-MY3 and MOST 104-2321-B-007-004. We are grateful for the use of the Tecnai F20 in the Cryo-EM Core Facility, Department of Academic Affairs and Instrument Service at Academia Sinica. Approved Most recent IF: 4.259
Call Number c:irua:134038 Serial 4087
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