Journal
JOURNAL OF STRUCTURAL BIOLOGY
Volume 181, Issue 2, Pages 136-148Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jsb.2012.12.006
Keywords
Contrast transfer function; Power Spectrum; Single particle analysis
Funding
- Spanish Ministry of Economy and Competitiveness [AIC-A-2011-0638, BFU2009-09331, BIO2010-16566, ACI2009-1022, ACI2010-1088, CAM(S2010/BMD-2305)]
- NSF [1114901]
- postdoctoral 'Juan de la Cierva Grants [JCI-2011-10185, JCI-2010-07594]
- Division Of Mathematical Sciences
- Direct For Mathematical & Physical Scien [1114901] Funding Source: National Science Foundation
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In this work we present a fast and automated algorithm for estimating the contrast transfer function (CTF) of a transmission electron microscope. The approach is very suitable for High Throughput work because: (a) it does not require any initial defocus estimation, (b) it is almost an order of magnitude faster than existing approaches, (c) it opens the way to well-defined extensions to the estimation of higher order aberrations, at the same time that provides defocus and astigmatism estimations comparable in accuracy to well established methods, such as Xmipp and CTFFIND3 approaches. The new algorithm is based on obtaining the wrapped modulating phase of the power spectra density pattern by the use of a quadrature filter. This phase is further unwrapped in order to obtain the continuous and smooth absolute phase map; then a Zernike polynomial fitting is performed and the defocus and astigmatism parameters are determined. While the method does not require an initial estimation of the defocus parameters or any non-linear optimization procedure, these approaches can be used if further refinement is desired. Results of the CTF estimation method are presented for standard negative stained images, cryo-electron microscopy images in the absence of carbon support, as well as micrographs with only ice. Additionally, we have also tested the proposed method with micrographs acquired from tilted and untilted samples, obtaining good results. The algorithm is freely available as a part of the Xmipp package [http:// xmipp.cnb.csic.es]. (C) 2012 Elsevier Inc. All rights reserved.
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