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From this figure, we have calculated an elongation percentage due to the missing wedge of 1.14%.
Images were taken from −76 ° to + 70 ° at 2 ° increments, resulting in a missing wedge of 34 °.
Electron tomographic reconstructions often contain artefacts from sources such as noise in the projections and a "missing wedge" of projection angles which can hamper quantitative analysis.
Tomography is closely related to CS, and even more so in electron tomography where it is common to have a missing wedge of data due to the inability to acquire all of the projections.
Unless tilt series are taken of pillar samples or individual particles and using a sample holder which allows full rotation without obstructions, there will always be a missing wedge of information for which to account.
To demonstrate the effect of the missing wedge, we use an angular range of −76 °– + 76 ° (and therefore a missing wedge of 28 °), which corresponds approximately to the maximum usable angular range for most tomographic experiments (unless free-standing needle samples are used).
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A dynamic, resolution-dependent, per-volume thresholding method was recently introduced (Galaz-Montoya et al. 2016a), which produces much more robust identification of the missing wedge than the use of a single threshold value, and better alignment results than adaptive low-pass filtering using estimates of the resolution (Hrabe et al. 2012) or FSC curves directly (Galaz-Montoya et al. 2015).
One caveat inherent to the thresholding method is that simple operations such as filtration and masking can alter the values of voxels inside or outside the missing wedge in different ways, leading to misidentification of missing wedge voxels and therefore increasing subtomogram alignment error.
The lack of detail in the membrane region is unlikely to be caused by the missing wedge as we have observed a similar phenomenon in sub-tomogram averages of other membrane proteins, which have no missing wedge of information, and in single-particle maps when the resolution does not extend beyond 15 Å.
Hence, the finding of complete rings is supported by the fact that perfect coincidence of any hypothetical gaps in constricting rings with the missing wedges in all of the analysed tomograms, found after all in entirely random orientations, would be remarkably implausible.
DOI: http://dx.doi.org/10.7554/eLife.03600.008 The orientation of our crystals prevented the full sampling of reciprocal space leading to systematic incompleteness (missing wedge or missing cone).
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