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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

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 cryog...

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Autores principales: Kobayashi, Amane, Sekiguchi, Yuki, Oroguchi, Tomotaka, Okajima, Koji, Fukuda, Asahi, Oide, Mao, Yamamoto, Masaki, Nakasako, Masayoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5357008/
https://www.ncbi.nlm.nih.gov/pubmed/27359147
http://dx.doi.org/10.1107/S1600577516007736
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author Kobayashi, Amane
Sekiguchi, Yuki
Oroguchi, Tomotaka
Okajima, Koji
Fukuda, Asahi
Oide, Mao
Yamamoto, Masaki
Nakasako, Masayoshi
author_facet Kobayashi, Amane
Sekiguchi, Yuki
Oroguchi, Tomotaka
Okajima, Koji
Fukuda, Asahi
Oide, Mao
Yamamoto, Masaki
Nakasako, Masayoshi
author_sort Kobayashi, Amane
collection PubMed
description 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.
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spelling pubmed-53570082017-03-24 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 J Synchrotron Radiat Research Papers 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. International Union of Crystallography 2016-05-31 /pmc/articles/PMC5357008/ /pubmed/27359147 http://dx.doi.org/10.1107/S1600577516007736 Text en © Amane Kobayashi et al. 2016 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.http://creativecommons.org/licenses/by/2.0/uk/
spellingShingle Research Papers
Kobayashi, Amane
Sekiguchi, Yuki
Oroguchi, Tomotaka
Okajima, Koji
Fukuda, Asahi
Oide, Mao
Yamamoto, Masaki
Nakasako, Masayoshi
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
title 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
title_full 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
title_fullStr 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
title_full_unstemmed 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
title_short 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
title_sort 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
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5357008/
https://www.ncbi.nlm.nih.gov/pubmed/27359147
http://dx.doi.org/10.1107/S1600577516007736
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