TY - JOUR
T1 - Diffraction apparatus and procedure in tomography X-ray diffraction imaging for biological cells at cryogenic temperature using synchrotron X-ray radiation
AU - Kobayashi, Amane
AU - Takayama, Yuki
AU - Okajima, Koji
AU - Oide, Mao
AU - Yamamoto, Takahiro
AU - Sekiguchi, Yuki
AU - Oroguchi, Tomotaka
AU - Nakasako, Masayoshi
AU - Kohmura, Yoshiki
AU - Yamamoto, Masaki
AU - Hoshi, Takahiko
AU - Torizuka, Yasufumi
N1 - Funding Information:
This study was supported by a grant for XFEL key technology, and by the X-ray Free Electron Laser Priority Strategy Program from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) to MN and MY. In addition, specimen preparation and software development were supported by grants from the Japan Society for the Promotion of Science to MN (jp17654084, jp23120525, jp24654140, jp25120725, jp16H02218), to YS (No. jp15J01707) and to AK (No. jp15J01831), and grants from the MEXT to MN (Nos. jp15076210, jp20050030).
Publisher Copyright:
© International Union of Crystallography, 2018
PY - 2018/11
Y1 - 2018/11
N2 - X-ray diffraction imaging is a technique for visualizing the structure of biological cells. In X-ray diffraction imaging experiments using synchrotron radiation, cryogenic conditions are necessary in order to reduce radiation damage in the biological cells. Frozen-hydrated biological specimens kept at cryogenic temperatures are also free from drying and bubbling, which occurs in wet specimens under vacuum conditions. In a previous study, the diffraction apparatus KOTOBUKI-1 [Nakasako et al. (2013), Rev. Sci. Instrum. 84, 093705] was constructed for X-ray diffraction imaging at cryogenic temperatures by utilizing a cryogenic pot, which is a cooling device developed in low-temperature physics. In this study a new cryogenic pot, suitable for tomography experiments, has been developed. The pot can rotate a biological cell over an angular range of ±170° against the direction of the incident X-ray beam. Herein, the details and the performance of the pot and miscellaneous devices are reported, along with established experimental procedures including specimen preparation. The apparatus has been used in tomography experiments for visualizing the three-dimensional structure of a Cyanidioschyzon merolae cell with an approximate size of 5 µm at a resolution of 136 nm. Based on the experimental results, the necessary improvements for future experiments and the resolution limit achievable under experimental conditions within a maximum tolerable dose are discussed.
AB - X-ray diffraction imaging is a technique for visualizing the structure of biological cells. In X-ray diffraction imaging experiments using synchrotron radiation, cryogenic conditions are necessary in order to reduce radiation damage in the biological cells. Frozen-hydrated biological specimens kept at cryogenic temperatures are also free from drying and bubbling, which occurs in wet specimens under vacuum conditions. In a previous study, the diffraction apparatus KOTOBUKI-1 [Nakasako et al. (2013), Rev. Sci. Instrum. 84, 093705] was constructed for X-ray diffraction imaging at cryogenic temperatures by utilizing a cryogenic pot, which is a cooling device developed in low-temperature physics. In this study a new cryogenic pot, suitable for tomography experiments, has been developed. The pot can rotate a biological cell over an angular range of ±170° against the direction of the incident X-ray beam. Herein, the details and the performance of the pot and miscellaneous devices are reported, along with established experimental procedures including specimen preparation. The apparatus has been used in tomography experiments for visualizing the three-dimensional structure of a Cyanidioschyzon merolae cell with an approximate size of 5 µm at a resolution of 136 nm. Based on the experimental results, the necessary improvements for future experiments and the resolution limit achievable under experimental conditions within a maximum tolerable dose are discussed.
KW - X-ray diffraction imaging
KW - biological cells
KW - cryogenic experiments
KW - tomography
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U2 - 10.1107/S1600577518012687
DO - 10.1107/S1600577518012687
M3 - Article
C2 - 30407193
AN - SCOPUS:85055322378
SN - 0909-0495
VL - 25
SP - 1803
EP - 1818
JO - Journal of Synchrotron Radiation
JF - Journal of Synchrotron Radiation
IS - 6
ER -