Combined Scanning Transmission Electron Microscopy Tilt- and Focal Series

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 a...

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Bibliographic Details
Published in:Microscopy and microanalysis 2014-04, Vol.20 (2), p.548-560
Main Authors: Dahmen, Tim, Baudoin, Jean-Pierre, Lupini, Andrew R., Kübel, Christian, Slusallek, Philipp, de Jonge, Niels
Format: Article
Language:English
Subjects:
Algorithms
Biological Applications
Carbon
Electron Microscope Tomography - methods
Electron microscopes
Imaging, Three-Dimensional - methods
Macrophages - ultrastructure
Microscopy, Electron, Scanning Transmission - methods
Models, Theoretical
Nanoparticles
Nanoparticles - ultrastructure
Silicon nitride
Software
Tomography
Transmission electron microscopy
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Summary: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.
ISSN:1431-9276
1435-8115
DOI:10.1017/S1431927614000075