Title
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Hollow cone electron imaging for single particle 3D reconstruction of proteins
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Author
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Abstract
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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 Å resolution but using a strongly reduced number of images. |
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Language
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English
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Source (journal)
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Scientific reports. - London, 2011, currens
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Publication
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London
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Nature Publishing Group
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2016
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ISSN
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2045-2322
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DOI
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10.1038/SREP27701
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Volume/pages
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6
(2016)
, p. 1-9
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Article Reference
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27701
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ISI
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000377670500001
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Medium
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E-only publicatie
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Full text (Publisher's DOI)
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Full text (open access)
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