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. Author manuscript; available in PMC: 2013 Feb 7.
Published in final edited form as: Nat Protoc. 2011 Aug 25;6(9):1391–1411. doi: 10.1038/nprot.2011.389

Figure 1.

Figure 1

The final high-resolution image obtained by our methods results from a two-step improvement of the PSF by deconvolution followed by Z-smear correction. A. The lateral and axial views of a dendritic spine in a raw confocal image, after deconvolution and after the Z-smear correction. Note that while the deconvolution improves both the lateral as well as the axial resolution, the Z-smear correction is exclusively aimed at the axial distortion that remains even after optimal deconvolution. B. The two-step improvement in the 3D resolution is plotted for 10 spines (various colors) and their group average (black line). There is a 49% reduction in volume by deconvolution and a further 40% improvement by the Z-smear correction (paired t-test, p<0.001 for each step). Put another way, a spine in a raw confocal stack is on average 226% bigger than its actual volume (range 77–294% for the 10 spines shown here). Animal use in this experiment was conducted with strict adherence to our Institutional Animal Care and Use Committee.