Correction for non-perpendicularity of beam and tilt axis in tomographic reconstructions with the IMOD package.
A correction for non-perpendicularity between the beam axis and the tilt axis in electron tomographic tilt series has been implemented in the IMOD software package and its value and limitations have been explored. Correction for this effect can provide a significant improvement in the alignment error and the reconstruction quality in some cases. However, when the projection model being fit includes an anisotropic shrinkage (i.e. stretch) in the plane of the specimen, adding a variable for the beam tilt does not produce a lower alignment error; it is thus not possible to distinguish between the effects of stretch and beam tilt. Test reconstructions indicate that an alignment solution that includes stretch will adequately correct for the effects of a beam tilt. For specimens subject to deformation under the beam, an alignment solution that accounts for stretch is preferable to one that accounts for beam tilt instead, provided that the markers used for alignment are sufficiently well distributed. Otherwise, a correction for beam tilt should be used.
Mastronarde
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DN
Boulder Laboratory for Three-dimensional Electron Microscopy of Cells, Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309, USA. mast@colorado.edu
D007089:Image Enhancement; D007090:Image Interpretation, Computer-Assisted; D008854:Microscopy, Electron / Q000379:methods*; D008954:Models, Biological; D012984:Software; D014054:Tomography / Q000379:methods*