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Record |
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Author |
Tsai, C.-Y.; Chang, Y.-C.; Lobato, I.; Van Dyck, D.; Chen, F.-R. |
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Title |
Hollow Cone Electron Imaging for Single Particle 3D Reconstruction of Proteins |
Type |
A1 Journal article |
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Year  |
2016 |
Publication |
Scientific reports |
Abbreviated Journal |
Sci Rep-Uk |
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Volume |
6 |
Issue |
6 |
Pages |
27701 |
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Keywords |
A1 Journal article; Electron microscopy for materials research (EMAT); Vision lab |
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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. |
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Address |
Department of Engineering and System Science, Tsing-Hua University, HsinChu 300, Taiwan |
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Corporate Author |
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Thesis |
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Publisher |
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Place of Publication |
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Editor |
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Language |
English |
Wos |
000377670500001 |
Publication Date |
2016-06-13 |
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Series Editor |
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Series Title |
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Abbreviated Series Title |
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Series Volume |
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Series Issue |
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Edition |
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ISSN |
2045-2322 |
ISBN |
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Additional Links |
UA library record; WoS full record; WoS citing articles |
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Impact Factor |
4.259 |
Times cited |
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Open Access |
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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 |
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Call Number |
c:irua:134038 |
Serial |
4087 |
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| Permanent link to this record |
