Resolution measures in molecular electron microscopy

By Pawel A. Penczek. 2010

138 citations recorded by the source corpus.

Resolution measures in molecular electron microscopy provide means to evaluate quality of macromolecular structures computed from sets of their two-dimensional line projections. When the amount of detail in the computed density map is low there are no external standards by which the resolution of the result can be judged. Instead, resolution measures in molecular electron microscopy evaluate consistency of the results in reciprocal space and present it as a one-dimensional function of the modulus of spatial frequency. Here we provide description of standard resolution measures commonly used in electron microscopy. We point out that the organizing principle is the relationship between these measures and the Spectral Signal-to-Noise Ratio of the computed density map. Within this framework it becomes straightforward to describe the connection between the outcome of resolution evaluations and the quality of electron microscopy maps, in particular, the optimum filtration, in the Wiener sense, of the computed map. We also provide a discussion of practical difficulties of evaluation of resolution in electron microscopy, particularly in terms of its sensitivity to data processing operations used during structure determination process in single particle analysis and in electron tomography.

Subfields: Materials Chemistry, Structural Biology, Surfaces, Coatings and Films

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Citations in the corpus

Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix (2019, background): “The overall resolution dFSC of a cryo-EM map is usually defined as the maximum spatial frequency at which the information content of the map is reliable (Penczek, 2010).”
A Primer to Single-Particle Cryo-Electron Microscopy (2015, unknown): “Once the CTF parameters have been determined and as long as a set of particle views that differ by defocus settings is available, correction for the CTF effects is possible and straightforward (Penczek, 2010a).”
New tools for the analysis and validation of cryo-EM maps and atomic models (2018, background): “Secondly, in practice some masking is typically applied to the map before Fourier coefficient calculation and this has a significant impact on the resulting values (Penczek, 2010; Pintilie et al., 2016).”
High-resolution noise substitution to measure overfitting and validate resolution in 3D structure determination by single particle electron cryomicroscopy (2013, background): “A number of interesting but pathological FSC curves, which illustrate some of the above points, are shown in Penczek's review on resolution [35].”
ATSAS 2.8: a comprehensive data analysis suite for small-angle scattering from macromolecular solutions (2017, methodology): “This approach is widely used in EM (Penczek, 2010), where the FSC is computed between the scattering amplitudes of the reconstructions based on independent experimental sets.”