Quantifying resolution limiting factors in subtomogram averaged cryo-electron tomography using simulations.

Cryo-electron tomography (CET) is the only available technique capable of characterizing the structure of biological macromolecules in conditions close to the native state. With the advent of subtomogram averaging, as a post-processing step to CET, resolutions in the (sub-) nanometer range have become within reach. In addition to advances in instrumentation and experiments, the reconstruction scheme has improved by inclusion of more accurate contrast transfer function (CTF) correction methods, better defocus estimation, and better alignments of the tilt-series and subtomograms.

Fast, spatially varying CTF correction in TEM.

We have developed new methods for contrast transfer function (CTF) correction of tilted and/or thick specimens. In order to achieve higher resolutions in cryo-electron tomography (CryoET), it is necessary to account for the defocus gradient on a tilted specimen and possibly the defocus gradient within a thick specimen. CTF correction methods which account for these defocus differences have recently gained interest.