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Nanoscale 3D cellular imaging by axial scanning transmission electron tomography

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Abstract

Electron tomography provides three-dimensional structural information about supramolecular assemblies and organelles in a cellular context, but image degradation, caused by scattering of transmitted electrons, limits applicability in specimens thicker than 300 nm. We found that scanning transmission electron tomography of 1,000-nm-thick samples using axial detection provided resolution comparable to that of conventional electron tomography. We demonstrated the method by reconstructing a human erythrocyte infected with the malaria parasite Plasmodium falciparum.

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Figure 1: STEM using axial detection.
Figure 2: Bright-field STEM analysis of 1-μm-thick C. reinhardtii.
Figure 3: The 3D ultrastructure of a human erythrocyte infected with P. falciparum.

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Acknowledgements

This work was supported by the Intramural Research Programs of the National Institute of Biomedical Imaging and Bioengineering, and the Eunice Kennedy Shriver National Institute of Child Health and Human Development of the National Institutes of Health. We thank T. Reese for help with the freeze-substitution technique. M.F.H.-M. acknowledges support through the Joint National Institute of Standards and Technology–National Institute of Biomedical Imaging and Bioengineering Postdoctoral Associateship program of the US National Research Council.

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Author notes

  1. Martin F Hohmann-Marriott

    Present address: Present address: University of Otago, Department of Biochemistry, Dunedin, New Zealand.,

  2. Martin F Hohmann-Marriott and Alioscka A Sousa: These authors contributed equally to this work.

Authors and Affiliations

  1. National Institute of Biomedical Imaging and Bioengineering, Bethesda, Maryland, USA

    Martin F Hohmann-Marriott, Alioscka A Sousa, Afrouz A Azari, Guofeng Zhang & Richard D Leapman

  2. Program in Physical Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA

    Svetlana Glushakova & Joshua Zimmerberg

Authors
  1. Martin F Hohmann-Marriott
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  2. Alioscka A Sousa
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  3. Afrouz A Azari
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  4. Svetlana Glushakova
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  5. Guofeng Zhang
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  6. Joshua Zimmerberg
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  7. Richard D Leapman
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Contributions

M.F.H.-M. and A.A.S. conceived the project, designed and performed experiments, processed and analyzed data and wrote the paper. A.A.A. processed and analyzed data. G.Z. prepared specimens for electron microscopy. S.G. and J.Z. provided samples of P. falciparum–infected erythrocytes, analyzed data and contributed to writing the paper. R.D.L. coordinated the project, designed experiments, analyzed data and wrote the paper.

Corresponding authors

Correspondence to Alioscka A Sousa or Richard D Leapman.

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Supplementary Text and Figures

Supplementary Figures 1–10 and Supplementary Discussion 1–2 (PDF 12187 kb)

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Hohmann-Marriott, M., Sousa, A., Azari, A. et al. Nanoscale 3D cellular imaging by axial scanning transmission electron tomography. Nat Methods 6, 729–731 (2009). https://doi.org/10.1038/nmeth.1367

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