Quantitative 3D imaging of whole, unstained cells by using X-ray diffraction microscopy

  • Huaidong Jiang
    Department of Physics and Astronomy and California NanoSystems Institute, University of California, Los Angeles, CA 90095;
  • Changyong Song
    RIKEN SPring-8 Center, 1-1-1, Kouto, Sayo, Hyogo 679-5148, Japan;
  • Chien-Chun Chen
    Department of Physics and Astronomy and California NanoSystems Institute, University of California, Los Angeles, CA 90095;
  • Rui Xu
    Department of Physics and Astronomy and California NanoSystems Institute, University of California, Los Angeles, CA 90095;
  • Kevin S. Raines
    Department of Physics and Astronomy and California NanoSystems Institute, University of California, Los Angeles, CA 90095;
  • Benjamin P. Fahimian
    Department of Physics and Astronomy and California NanoSystems Institute, University of California, Los Angeles, CA 90095;
  • Chien-Hung Lu
    Institute of Physics, Academia Sinica, Nankang, Taipei 11529, Taiwan; and
  • Ting-Kuo Lee
    Institute of Physics, Academia Sinica, Nankang, Taipei 11529, Taiwan; and
  • Akio Nakashima
    Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095
  • Jun Urano
    Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095
  • Tetsuya Ishikawa
    RIKEN SPring-8 Center, 1-1-1, Kouto, Sayo, Hyogo 679-5148, Japan;
  • Fuyuhiko Tamanoi
    Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095
  • Jianwei Miao
    Department of Physics and Astronomy and California NanoSystems Institute, University of California, Los Angeles, CA 90095;

書誌事項

公開日
2010-06-04
DOI
  • 10.1073/pnas.1000156107
公開者
Proceedings of the National Academy of Sciences

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説明

<jats:p>Microscopy has greatly advanced our understanding of biology. Although significant progress has recently been made in optical microscopy to break the diffraction-limit barrier, reliance of such techniques on fluorescent labeling technologies prohibits quantitative 3D imaging of the entire contents of cells. Cryoelectron microscopy can image pleomorphic structures at a resolution of 3–5 nm, but is only applicable to thin or sectioned specimens. Here, we report quantitative 3D imaging of a whole, unstained cell at a resolution of 50–60 nm by X-ray diffraction microscopy. We identified the 3D morphology and structure of cellular organelles including cell wall, vacuole, endoplasmic reticulum, mitochondria, granules, nucleus, and nucleolus inside a yeast spore cell. Furthermore, we observed a 3D structure protruding from the reconstructed yeast spore, suggesting the spore germination process. Using cryogenic technologies, a 3D resolution of 5–10 nm should be achievable by X-ray diffraction microscopy. This work hence paves a way for quantitative 3D imaging of a wide range of biological specimens at nanometer-scale resolutions that are too thick for electron microscopy.</jats:p>

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