4.8 Article

Three-dimensional cellular ultrastructure resolved by X-ray microscopy

Journal

NATURE METHODS
Volume 7, Issue 12, Pages 985-U116

Publisher

NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.1533

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Funding

  1. Human Frontier Science Program [RGP0053/2005-C]
  2. German Federal Ministry of Education and Research [05KS4BY1/7]
  3. US National Institutes of Health
  4. National Institute of Arthritis and Musculoskeletal and Skin Diseases, and the National Cancer Institute, Center for Cancer Research [HHSN26120080001E]

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We developed an X-ray microscope using partially coherent object illumination instead of previously used quasi-incoherent illumination. The design permitted the incorporation of a cryogenic tilt stage, enabling tomography of frozen-hydrated, intact adherent cells. We obtained three-dimensional reconstructions of mouse adenocarcinoma cells at similar to 36-nm (Rayleigh) and similar to 70-nm (Fourier ring correlation) resolution, which allowed us to visualize the double nuclear membrane, nuclear pores, nuclear membrane channels, mitochondrial cristae and lysosomal inclusions.

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