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Combined Scanning Transmission Electron Microscopy Tilt- and Focal Series

Published online by Cambridge University Press:  19 February 2014

Tim Dahmen
Affiliation:
German Research Center for Artificial Intelligence GmbH (DFKI), 66123 Saarbrücken, Germany
Jean-Pierre Baudoin
Affiliation:
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232-0615, USA
Andrew R. Lupini
Affiliation:
Karlsruhe Institute for Technology (KIT), 76344 Eggenstein-Leopoldshafen, Germany
Christian Kübel
Affiliation:
Karlsruhe Institute for Technology (KIT), 76344 Eggenstein-Leopoldshafen, Germany
Philipp Slusallek
Affiliation:
German Research Center for Artificial Intelligence GmbH (DFKI), 66123 Saarbrücken, Germany
Niels de Jonge
Affiliation:
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232-0615, USA Leibniz Institute for New Materials (INM), 66123 Saarbrücken, Germany
Corresponding
E-mail address:

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.

Type
Biological Applications
Copyright
© Microscopy Society of America 2014 

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Footnotes

Current address: La Timone Hospital and Medicine School, Marseille, France

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Dahmen Supplementary Material

Combined tilt- and focal series high annular dark field scanning transmission electron microscopy (STEM) data. The sample was a whole mount macrophage cell containing gold nanoparticles of two different sizes distributed in clusters throughout its volume. The movie represents the aligned data recorded for a tilt range of -40° to +40° in 5° increments, with a focal series at each tilt angle using focus steps of 50 nm.

Download Dahmen Supplementary Material(Video)
Video 9 MB

Dahmen Supplementary Material

Perspective rendering of the reconstructed tomogram of the same cellular region as shown in Movie S1. The signal intensity was color-coded. The tilt-focal algebraic reconstruction technique (TF-ART) was used for the three dimensional reconstruction.

Download Dahmen Supplementary Material(Video)
Video 23 MB
Supplementary material: File

Dahmen Supplementary Material

Movie Captions

Download Dahmen Supplementary Material(File)
File 24 KB
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