Reliable cryo-EM resolution estimation with modified Fourier shell correlation

A modified Fourier shell correlation (mFSC) methodology is introduced that is aimed at addressing two fundamental problems that mar the use of the FSC: the strong influence of mask-induced artifacts on resolution estimation and the lack of assessment of FSC uncertainties stemming from the inability...

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Bibliographic Details
Published in:IUCrJ 2020-11, Vol.7 (6), p.995-1008
Main Author: Penczek, Pawel A.
Format: Article
Language:English
Subjects:
Computation
Correlation
cryo-em
Degrees of freedom
Expected values
Gaussian window functions
modified fourier shell correlation
Noise
Research Papers
resolution
signal-to-noise ratio
Standard deviation
three-dimensional structure determination
Window functions
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Summary:A modified Fourier shell correlation (mFSC) methodology is introduced that is aimed at addressing two fundamental problems that mar the use of the FSC: the strong influence of mask-induced artifacts on resolution estimation and the lack of assessment of FSC uncertainties stemming from the inability to determine the associated number of degrees of freedom. It is shown that by simply changing the order of the steps in which the FSC is computed, the correlations induced by masking of the input data can be eliminated. In addition, to further reduce artifacts, a smooth Gaussian window function is used to outline the regions of reciprocal space within which the mFSC is computed. Next, it is shown that the number of degrees of freedom (ndf) of the system is approximated well by combining the ndf associated with the Gaussian window in reciprocal space with further reduction of the ndf owing to the use of the mask in real space. It is demonstrated through the application of the mFSC to both single-particle and helical structures that the mFSC yields reliable, mask-induced artifact-free results as a result of the introduced modifications. Since the adverse effect of the mask is eliminated, it also becomes possible to compute robust local resolutions both per voxel of a 3D map as well as, in a newly developed approach, per functional subunit, segment or even larger secondary element of the studied complex.
ISSN:2052-2525
2052-2525
DOI:10.1107/S2052252520011574