Quantitative 3D imaging of whole, unstained cells by using X-ray diffraction microscopy
| Autor: | Tetsuya Ishikawa, Fuyuhiko Tamanoi, Jianwei Miao, Ting-Kuo Lee, Kevin S. Raines, Akio Nakashima, Benjamin P. Fahimian, Chien-Hung Lu, Rui Xu, Jun Urano, Changyong Song, Huaidong Jiang, Chien-Chun Chen |
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| Rok vydání: | 2010 |
| Předmět: |
Optics and Photonics
Analytical chemistry Electrons Biology Models Biological law.invention Biological specimen Imaging Three-Dimensional Optical microscope X-Ray Diffraction law Yeasts Microscopy Schizosaccharomyces Spore germination Image Processing Computer-Assisted Scattering Radiation Multidisciplinary Models Statistical X-Rays Resolution (electron density) Cryoelectron Microscopy Scanning confocal electron microscopy Spores Fungal Dark field microscopy Physical Sciences Biophysics Electron microscope Algorithms |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 107(25) |
| ISSN: | 1091-6490 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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