Combined scanning transmission electron microscopy tilt- and focal series

Microsc Microanal. 2014 Apr;20(2):548-60. doi: 10.1017/S1431927614000075. Epub 2014 Feb 19.

Abstract

In this study, a combined tilt- and focal series is proposed as a new recording scheme for high-angle annular dark-field scanning transmission electron microscopy (STEM) tomography. Three-dimensional (3D) data were acquired by mechanically tilting the specimen, and recording a through-focal series at each tilt direction. The sample was a whole-mount macrophage cell with embedded gold nanoparticles. The tilt-focal algebraic reconstruction technique (TF-ART) is introduced as a new algorithm to reconstruct tomograms from such combined tilt- and focal series. The feasibility of TF-ART was demonstrated by 3D reconstruction of the experimental 3D data. The results were compared with a conventional STEM tilt series of a similar sample. The combined tilt- and focal series led to smaller "missing wedge" artifacts, and a higher axial resolution than obtained for the STEM tilt series, thus improving on one of the main issues of tilt series-based electron tomography.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Algorithms
  • Artifacts
  • Electron Microscope Tomography / methods*
  • Imaging, Three-Dimensional / methods
  • Macrophages / ultrastructure
  • Microscopy, Electron, Scanning Transmission / methods*
  • Models, Theoretical
  • Nanoparticles / ultrastructure