抄録
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.
| 元の言語 | English |
|---|---|
| ページ（範囲） | 975-989 |
| ページ数 | 15 |
| ジャーナル | Journal of Synchrotron Radiation |
| 巻 | 23 |
| 発行部数 | 4 |
| DOI | |
| 出版物ステータス | Published - 2016 7 1 |
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ASJC Scopus subject areas
- Instrumentation
- Nuclear and High Energy Physics
- Radiation
これを引用
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. / Kobayashi, Amane; Sekiguchi, Yuki; Oroguchi, Tomotaka; Okajima, Koji; Fukuda, Asahi; Oide, Mao; Yamamoto, Masaki; Nakasako, Masayoshi.
：: Journal of Synchrotron Radiation, 巻 23, 番号 4, 01.07.2016, p. 975-989.研究成果: Article
}
TY - JOUR
T1 - 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
AU - Kobayashi, Amane
AU - Sekiguchi, Yuki
AU - Oroguchi, Tomotaka
AU - Okajima, Koji
AU - Fukuda, Asahi
AU - Oide, Mao
AU - Yamamoto, Masaki
AU - Nakasako, Masayoshi
PY - 2016/7/1
Y1 - 2016/7/1
N2 - 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.
AB - 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.
KW - coherent X-ray diffraction imaging
KW - cryogenic X-ray diffraction experiment
KW - frozen-hydrated non-crystalline specimens
KW - structures of cells and cellular organelles
KW - X-ray free-electron laser
UR - http://www.scopus.com/inward/record.url?scp=84976647543&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84976647543&partnerID=8YFLogxK
U2 - 10.1107/S1600577516007736
DO - 10.1107/S1600577516007736
M3 - Article
C2 - 27359147
AN - SCOPUS:84976647543
VL - 23
SP - 975
EP - 989
JO - Journal of Synchrotron Radiation
JF - Journal of Synchrotron Radiation
SN - 0909-0495
IS - 4
ER -