Development of a hybrid microscope to observe inner structures and their variation of living biological cells in situ.

• Project/Area Number: 25390134
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Quantum beam science
• Research Institution: Japan Atomic Energy Agency
• Principal Investigator: KADO Masataka 国立研究開発法人日本原子力研究開発機構, 原子力科学研究部門 量子ビーム応用研究センター, 研究主幹 (30360431)
• Co-Investigator(Renkei-kenkyūsha): KISHIMOTO Maki 国立研究開発法人日本原子力研究開発機構, 原子力科学研究部門 量子ビーム応用研究センター, 研究主幹 (40360432)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000); Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 軟X線顕微鏡 / 相関顕微法 / 細胞内小器官 / アポトーシス / 生きている細胞 / レーザープラズマ軟X線顕微鏡 / 蛍光顕微鏡 / ハイブリッド顕微鏡 / 生きている細胞の観察 / ミトコンドリア

Outline of Final Research Achievements

I have developed a hybrid microscopy which is a combination of a fluorescent microscope and a soft X-ray microscope and this microscopy enabled to observe live biological cells with both microscopes at the same time. Observing Leydig cells of mouse testis with the hybrid microscopy, I have succeeded to obtain a soft X-ray image of a single mitochondrion in a live biological cell. The hybrid microscopy also was applied to observe apoptotic cells and details of structural deformation due to the apoptosis have been cleared.

Research Products

• [Journal Article] Correlative Imaging of Live Biological Cells with a Soft X-ray Microscope and a Fluorescence Microscope (2016)
  • Author(s): M. Kado, M. Kishimoto, S. Tamotsu, K. Yasuda, M. Aoyama, S. Tone and K. Shinohara
  • Journal Title: AIP Conf. Proc. Volume: 1696 Pages: 020019-020019
  • DOI: 10.1063/1.4937513
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] In situ observation of cellular organelles with a contact x-ray microscop (2013)
  • Author(s): M Kado, M Kishimoto, S Tamotsu, K Yasuda and K Shinohara
  • Journal Title: Journal ofPhysics:ConferenceSeries Volume: 463 Pages: 12056-12056
  • DOI: 10.1088/1742-6596/463/1/012056
  • Peer Reviewed
• [Journal Article] Imaging of fine structures of cellular organelles in hydrated biological cells by a soft x-ray microscope combined with a fluorescence microscope (2013)
  • Author(s): M. Kado, M. Kishimoto, S. Tamotsu, K. Yasuda, M. Aoyama, and K. Shinohara
  • Journal Title: Proc. of SPIE Volume: 8849 Pages: 88490C-88490C
  • DOI: 10.1117/12.2022332
• [Presentation] レーザープラズマ軟X線顕微鏡による生きている細胞の内部構 造の観察 (2016)
  • Author(s): 加道雅孝
  • Organizer: 「プラズマ科学における分光計測の高度化と原子分子過程研究の新展開」 「原子分子データ応用フォーラムセミナー」合同研究会
  • Place of Presentation: 核融合科学研究所（岐阜県土岐市）
  • Year and Date: 2016-01-28
  • Invited
• [Presentation] レーザープラズマ軟X線顕微鏡を用いた相関顕微法による細胞内小器官の観察 (2016)
  • Author(s): 加道雅孝，岸本牧，保智己 ，安田恵子，青山雅人，刀祢重信，篠原邦夫
  • Organizer: レーザー学会学術講演会第36回年次大会
  • Place of Presentation: 名城大学天白キャンパス（愛知県名古屋市）
  • Year and Date: 2016-01-10
  • Invited
• [Presentation] 高輝度レーザープラズマ軟X線源を用いたアポトーシス誘発細胞核の軟X線顕微鏡観察 (2015)
  • Author(s): 加道雅孝、岸本牧、刀祢重信、保智己、安田恵子、青山雅人、篠原邦夫
  • Organizer: 第38 回日本分子生物学会年会
  • Place of Presentation: 神戸国際会議場（兵庫県神戸市）
  • Year and Date: 2015-12-04
• [Presentation] Correlative microscopy with a laser-plasma soft X-ray microscope and a fluorescent microscope for biological imaging of live hydrated cells (2015)
  • Author(s): Kado M., Kishimoto M., Tamotsu S., Yasuda K., Aoyama M., Tone S., Shinohara K.
  • Organizer: Microscopy Conference 2015
  • Place of Presentation: ゲッチンゲン（ドイツ）
  • Year and Date: 2015-09-06
  • Int'l Joint Research
• [Presentation] Live cell imaging with a soft X-ray microscope using a laser-plasma soft X-ray source (2015)
  • Author(s): Masataka Kado, Maki Kishimoto, Masahiko Ishino, Satoshi Tamotsu, Keiko Yasuda, Masato Aoyama, Shigenobu Tone and Kunio Shinohara
  • Organizer: 1st Workshop on Applications of Laser Plasma X-ray and EUV Sources in Technology and Science (ALPS 2015)
  • Place of Presentation: ワルシャワ（ポーランド）
  • Year and Date: 2015-07-06
  • Int'l Joint Research / Invited
• [Presentation] Development of a soft x-ray microscope with a intense laser-plasma soft x-ray source produced by a high power pulsed laser (2015)
  • Author(s): M. Kado, M. Kishimoto, M. Ishino, S. Tamotsu, K. Yasuda, M. Aoyama, S. Tone, K. Shinohara
  • Organizer: HEDS in Asia 2014
  • Place of Presentation: ロナワラ（インド）
  • Year and Date: 2015-01-20 – 2015-01-21
  • Invited
• [Presentation] レーザープラズマ軟X 線顕微鏡による生きている細胞の内部構造の観察 (2014)
  • Author(s): 加道雅孝
  • Organizer: 平成26 年第1 回「顕微表面・界面分析セミナー
  • Place of Presentation: 島津製作所関西支社マルチホール（大阪市）
  • Year and Date: 2014-12-19
  • Invited
• [Presentation] 軟X 線顕微鏡による細胞内部構造のその場観察 (2014)
  • Author(s): 加道雅孝、岸本牧、保智己、安田恵子、青山雅人、刀祢重信、篠原邦夫
  • Organizer: 第4 回宇都宮大学オプトバイオシンポジウム
  • Place of Presentation: 宇都宮大学（宇都宮市）
  • Year and Date: 2014-12-11
  • Invited
• [Presentation] 生きている細胞の内部構造を直接観察できる軟X 線顕微鏡の開発 (2014)
  • Author(s): 加道雅孝、岸本牧、保智己、安田恵子、青山雅人、篠原邦夫
  • Organizer: 第37 回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（横浜市）
  • Year and Date: 2014-11-25 – 2014-11-28
• [Presentation] レーザー生成プラズマ軟X 線源の高輝度化と軟X 線顕微鏡による生きている細胞の観察 (2014)
  • Author(s): 加道雅孝、岸本牧、江島丈雄、保智己、安田恵子、青山雅人、篠原邦夫
  • Organizer: Plasma Conference 2014
  • Place of Presentation: 朱鷺メッセ（新潟市）
  • Year and Date: 2014-11-18 – 2014-11-21
• [Presentation] Hybrid imaging of living biological cells with a soft X-ray microscope and a fluorescent microscope (2014)
  • Author(s): M. Kado, M. Kishimoto, S. Tamotsu, K. Yasuda, M. Aoyama, S. Tone, K. Shinohara
  • Organizer: International Conference on X-ray Microscopy
  • Place of Presentation: メルボルン（オーストラリア）
  • Year and Date: 2014-10-27 – 2014-10-31
• [Presentation] 試料周辺の水の層の厚さの制御による密着型軟X 線顕微鏡の高コントラスト化 (2014)
  • Author(s): 加道雅孝、岸本牧、刀祢重信、保智己、安田恵子、青山雅人、篠原邦夫
  • Organizer: 第７５ 回応用物理学会秋季学術講演会
  • Place of Presentation: 北海道大学（札幌市）
  • Year and Date: 2014-09-17 – 2014-09-20
• [Presentation] Single flash imaging of live hydrated biological cells by a contact soft x-ray microscope coupled with an intense laser-plasma soft x-ray source (2014)
  • Author(s): M. Kado, M. Kishimoto, S. Tamotsu, K. Yasuda, M. Aoyama, K. Shinohara
  • Organizer: 18th International Microscopy Congress
  • Place of Presentation: プラハ（チェコ）
  • Year and Date: 2014-09-08 – 2014-09-12
• [Presentation] Soft X-ray emissions from ultra-thin foiled targets irradiated with an intense pulsed laser (2014)
  • Author(s): M. Kado, M. Ishino, M. Kishimoto, S. Tamotsu, K. Yasuda, K. Shinohara
  • Organizer: 33rd European Conference on Laser Interaction with Matter
  • Place of Presentation: パリ（フランス）
  • Year and Date: 2014-09-01 – 2014-09-05
• [Presentation] Development of a soft x-ray microscope with intense laser-plasma soft x-ray source produced by a high power pulsed laser (2014)
  • Author(s): Masataka Kado, Maki Kishimoto, Masahiko Ishino, Satoshi Tamotsu, Keiko Yasuda, Masato Aoyama, Shigenobu Tone and Kunio Shinohara
  • Organizer: HEDS in Asia 2014
  • Place of Presentation: 釜山(韓国)
  • Invited
• [Presentation] Structural Study of Cellular Organelles with a Laser-Plasma Soft X-Ray Microscope (2013)
  • Author(s): Masataka Kado, Maki Kishimoto, Satoshi Tamotsu, Keiko Yasuda, Masato Aoyama and Kunio Shinohara
  • Organizer: The 12th Symposium on X-ray Imaging Optics
  • Place of Presentation: 大阪市
  • Invited
• [Presentation] Imaging of fine structures of cellular organelles in hydrated biological cells by a soft x-ray microscope combined with a fluorescence microscope (2013)
  • Author(s): M. Kado, M. Kishimoto, S. Tamotsu, K. Yasuda, M. Aoyama, and K. Shinohara
  • Organizer: SPIE 2013
  • Place of Presentation: サンディエゴ(米国)
• [Presentation] レーザープラズマ軟X線顕微鏡によるアポトーシス細胞核の観察II (2013)
  • Author(s): 加道雅孝、岸本牧、刀祢重信、保智己、安田恵子、青山雅人、篠原邦夫
  • Organizer: 第74 回応用物理学会秋季学術講演会
  • Place of Presentation: 京田辺市
• [Presentation] レーザープラズマ軟X線顕微鏡によるアポトーシス細胞核の観察 (2013)
  • Author(s): 加道雅孝、岸本牧、刀祢重信、保智己、安田恵子、青山雅人、篠原邦夫
  • Organizer: レーザー学会第455回研究会
  • Place of Presentation: 敦賀市