Specimen preparation for cryogenic coherent X-ray diffraction imaging of biological cells and cellular organelles by using the X-ray free-electron laser at SACLA

Amane Kobayashi, Yuki Sekiguchi, Tomotaka Oroguchi, Koji Okajima, Asahi Fukuda, Mao Oide, Masaki Yamamoto, Masayoshi Nakasako

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Coherent X-ray diffraction imaging (CXDI) allows internal structures of biological cells and cellular organelles to be analyzed. CXDI experiments have been conducted at 66 K for frozen-hydrated biological specimens at the SPring-8 Angstrom Compact Free-Electron Laser facility (SACLA). In these cryogenic CXDI experiments using X-ray free-electron laser (XFEL) pulses, specimen particles dispersed on thin membranes of specimen disks are transferred into the vacuum chamber of a diffraction apparatus. Because focused single XFEL pulses destroy specimen particles at the atomic level, diffraction patterns are collected through raster scanning the specimen disks to provide fresh specimen particles in the irradiation area. The efficiency of diffraction data collection in cryogenic experiments depends on the quality of the prepared specimens. Here, detailed procedures for preparing frozen-hydrated biological specimens, particularly thin membranes and devices developed in our laboratory, are reported. In addition, the quality of the frozen-hydrated specimens are evaluated by analyzing the characteristics of the collected diffraction patterns. Based on the experimental results, the internal structures of the frozen-hydrated specimens and the future development for efficient diffraction data collection are discussed.Detailed procedures and key points in preparing frozen-hydrated biological specimens are reported for efficient cryogenic coherent X-ray diffraction imaging using an X-ray free-electron laser.

Original languageEnglish
Pages (from-to)975-989
Number of pages15
JournalJournal of Synchrotron Radiation
Volume23
Issue number4
DOIs
Publication statusPublished - 2016 Jul 1

Keywords

  • X-ray free-electron laser
  • coherent X-ray diffraction imaging
  • cryogenic X-ray diffraction experiment
  • frozen-hydrated non-crystalline specimens
  • structures of cells and cellular organelles

ASJC Scopus subject areas

  • Radiation
  • Nuclear and High Energy Physics
  • Instrumentation

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