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J Struct Biol. 2015 Nov;192(2):297-306. doi: 10.1016/j.jsb.2015.10.004. Epub 2015 Oct 9.

A fast cross-validation method for alignment of electron tomography images based on Beer-Lambert law.

Author information

1
Markey Center for Structural Biology, Department of Biological Sciences, Purdue University, West Lafayette, IN 47907, USA.
2
Department of Pathology and Laboratory Medicine, University of Texas Medical School at Houston, Houston, TX 77030, USA.
3
Markey Center for Structural Biology, Department of Biological Sciences, Purdue University, West Lafayette, IN 47907, USA. Electronic address: jiang12@purdue.edu.

Abstract

In electron tomography, accurate alignment of tilt series is an essential step in attaining high-resolution 3D reconstructions. Nevertheless, quantitative assessment of alignment quality has remained a challenging issue, even though many alignment methods have been reported. Here, we report a fast and accurate method, tomoAlignEval, based on the Beer-Lambert law, for the evaluation of alignment quality. Our method is able to globally estimate the alignment accuracy by measuring the goodness of log-linear relationship of the beam intensity attenuations at different tilt angles. Extensive tests with experimental data demonstrated its robust performance with stained and cryo samples. Our method is not only significantly faster but also more sensitive than measurements of tomogram resolution using Fourier shell correlation method (FSCe/o). From these tests, we also conclude that while current alignment methods are sufficiently accurate for stained samples, inaccurate alignments remain a major limitation for high resolution cryo-electron tomography.

KEYWORDS:

Alignment accuracy; Beer–Lambert law; Cross validation; Electron tomography; Inelastic scattering; Least square fitting

PMID:
26455556
PMCID:
PMC4633358
DOI:
10.1016/j.jsb.2015.10.004
[Indexed for MEDLINE]
Free PMC Article

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