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For this reason, we have represented the image in Fourier space using the real-space image as a proxy to facilitate interpretation of the impact of missing wedge bias.
Open image in new window Fig. 7 Missing wedge bias in Fourier space.
Preventing "missing wedge bias" is an essential step to accomplish this.
This "missing wedge bias" towards favoring larger density overlaps can cause misalignment of subtomograms; however, it can be compensated for by cross correlation map normalization or other methods.
To avoid missing wedge bias, we used cross-correlation map normalization (Schmid and Booth, 2008).
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We show that the missing wedge causes a bias in classification, and that this bias can be minimized by supplementation with data from the Fourier transform of the averaged structure.
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).
Fig. 5 Missing wedge reconstruction, processed with NAD. a A WBP reconstruction of aligned, noiseless projections with a 28 ° missing wedge.
Fig. 10 Missing wedge reconstruction processed with Weka. a The perfectly aligned, noiseless 28 ° missing wedge reconstruction from Fig. 5. b The same reconstruction but processed with Trainable Weka Segmentation.
Despite the recent progress in the field of reconstruction and regularization techniques, the missing wedge artefacts remain untreated in most current reconstruction workflows in structural biology.
Due to the missing wedge artifact, thylakoid grana that may have been present at the top and bottom of the chloroplast could not be resolved.
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