Coherent X-Ray Diffraction Imaging of Chloroplasts from Cyanidioschyzon merolae by Using X-Ray Free Electron Laser

Plant & Cell Physiology
Yuki TakayamaMasayoshi Nakasako

Abstract

Coherent X-ray diffraction imaging (CXDI) is a lens-less technique for visualizing the structures of non-crystalline particles with the dimensions of submicrometer to micrometer at a resolution of several tens of nanometers. We conducted cryogenic CXDI experiments at 66 K to visualize the internal structures of frozen-hydrated chloroplasts of Cyanidioschyzon merolae using X-ray free electron laser (XFEL) as a coherent X-ray source. Chloroplast dispersed specimen disks at a number density of 7/(10×10 µm(2)) were flash-cooled with liquid ethane without staining, sectioning or chemical labeling. Chloroplasts are destroyed at atomic level immediately after the diffraction by XFEL pulses. Thus, diffraction patterns with a good signal-to-noise ratio from single chloroplasts were selected from many diffraction patterns collected through scanning specimen disks to provide fresh specimens into the irradiation area. The electron density maps of single chloroplasts projected along the direction of the incident X-ray beam were reconstructed by using the iterative phase-retrieval method and multivariate analyses. The electron density map at a resolution of 70 nm appeared as a C-shape. In addition, the fluorescence image of proteins stained ...Continue Reading

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Citations

Feb 27, 2016·Acta Crystallographica. Section A, Foundations and Advances·Yuki Takayama, Koji Yonekura
Jan 18, 2018·Scientific Reports·Amane KobayashiMasayoshi Nakasako
Jul 1, 2017·Journal of Synchrotron Radiation·Miki NakanoFlorence Tama
Jul 7, 2018·Journal of Synchrotron Radiation·Yuki TakayamaYasushi Kagoshima
Nov 25, 2018·Optics Express·Tetsuro NagaiOsamu Miyashita
Jul 15, 2015·Plant & Cell Physiology·Akira Nagatani, Tetsuro Mimura
Mar 18, 2020·Biophysics Reviews·Masayoshi NakasakoMasaki Yamamoto

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