Three-dimensional super-resolution spectromicroscopy to trace inter-protein interactions and morphological changes in photosynthetic membranes on spatial and temporal domains

• Project Area: Molecular Science for Supra Functional Systems ? Development of Advanced Methods for Exploring Elementary Process
• Project/Area Number: 19056012
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Kyoto University
• Principal Investigator: KUMAZAKI Shigeichi 京都大学, 大学院・理学研究科, 准教授 (40293401)
• Project Period (FY): 2007 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥33,300,000 (Direct Cost: ¥33,300,000); Fiscal Year 2011: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 2010: ¥3,700,000 (Direct Cost: ¥3,700,000); Fiscal Year 2009: ¥9,700,000 (Direct Cost: ¥9,700,000); Fiscal Year 2008: ¥10,100,000 (Direct Cost: ¥10,100,000); Fiscal Year 2007: ¥8,200,000 (Direct Cost: ¥8,200,000)
• Keywords: シアノバクテリア / チラコイド膜 / 葉緑体 / 顕微分光 / 光化学系 / 2光子励起 / アンチストークス蛍光 / 近赤外レーザー / 蛍光スペクトル / 多光子励起 / 光化学系I / 光化学系II / ヘテロシスト / 蛍光顕微鏡 / クロロフィル / 植物 / 超解像 / 生物物理化学 / 電子・エネルギー移動 / 色素体機能・光合成 / 光計測 / 光合成 / 二光子励起 / 顕微蛍光 / 共焦点顕微鏡 / 緑藻

Research Abstract

We have found that photosystem I (PSI)-specific fluorescence spectra of thylakoid membranes of cyanobacteria and chloroplasts was very efficiently observed in the case of continuous wave (CW) near infrared (NIR) excitation around 785-820 nm. It was found that the PSI fluorescence in the case of CW NIR excitation is mainly generated by a one-photon excitation mechanism. This phenomenon will be used for studies of PSI/PSII ratio at physiological temperature, since PSI fluorescence has been conventionally observed at low temperatures like 77 K.

Research Products

• [Journal Article] Microspectroscopic Analysis of Photosynthetic Membrane by Anti-stokes Fluorescence and Two-photon Excitation Fluorescence (2011)
  • Author(s): 熊崎茂一
  • Journal Title: Seibutsu Butsuri Volume: 51 Issue: 6 Pages: 274-275
  • DOI: 10.2142/biophys.51.274
  • NAID: 10030038018
  • ISSN: 0582-4052, 1347-4219
• [Journal Article] Anti-Stokes Fluorescence Spectra of Chloroplasts in Parachlorella kessleri and Maize at Room Temperature as Characterized by Near Infrared Continuous Wave Laser Fluorescence Microscopy and Absorption Microscopy (2011)
  • Author(s): Makoto Hasegawa, Takahiko Yoshida, Mitsunori Yabuta, Masahide Terazima, and Shigeichi Kumazaki
  • Journal Title: J. Phys. Chem. B. Volume: 115 Issue: 14 Pages: 4184-4194
  • DOI: 10.1021/jp111306k
• [Journal Article] ライン走査型2光子励起蛍光スペクトル顕微鏡を用いた植物細胞中の葉緑体内部微細構造観察 (2011)
  • Author(s): 熊崎茂一, 長谷川慎
  • Journal Title: 分光研究 Volume: 60(1) Pages: 19-21
  • NAID: 10027959134
• [Journal Article] Anti-Stokes Fluorescence Spectra of Chloroplasts in Parachlorella kessleri and Maize at Room Temperature as Characterized by Near Infrared Continuous Wave Laser Fluorescence Microscopy and Absorption Microscopy (2011)
  • Author(s): Makoto Hasegawa, Takahiko Yoshida, Mitsunori Yabuta, Masahide Terazima, Shigeichi Kumazaki
  • Journal Title: Journal of Physical Chemistry B Volume: 115 Pages: 4184-4194
  • Peer Reviewed
• [Journal Article] Selective Excitation of Photosystems in Chloroplasts inside Plant Leaves Observed by Near-Infrared Laser-Based Fluorescence Spectral Microscopy (2010)
  • Author(s): Makoto Hasegawa, Takashi Shiina, Masahide Terazima and *Shigeichi Kumazaki
  • Journal Title: Plant Cell Physiol. Volume: 51 Issue: 2 Pages: 225-238
  • DOI: 10.1093/pcp/pcp182
  • NAID: 10027346696
  • Peer Reviewed
• [Journal Article] Selective Excitation of Photosystems in Chloroplasts inside Plant Leaves Observed by Near-Infrared Laser-Based Fluorescence Spectral Microscopy (2010)
  • Author(s): M. Hasegawa, T. Shiina, M. Terazima, S. Kumazaki
  • Journal Title: Plant Cell Physiology 51 Pages: 225-238
  • NAID: 10027346696
  • Peer Reviewed
• [Journal Article] A line-scanning semi-confocal multi-photon fluorescence microscope with a simultaneous broadband spectral acquisition and its application to the study of the thylakoid membrane of a cyanobacterium Anabaena PCC7120 (2007)
  • Author(s): Shigeichi. Kumazaki, Makoto Hasegawa, Mohammad Ghoneim, Yugo Shimizu, Kenji Okamoto, M. Nishiyama, H. Oh-oka and M. Terazima
  • Journal Title: J. Microsc. Volume: 228 Issue: 2 Pages: 240-254
  • DOI: 10.1111/j.1365-2818.2007.01835.x
• [Journal Article] A Line-Scanning Semiconfocal Multiphoton Fluorescence Microscope with a Simultaneous Broadband Spectral Acquisition and its Application to the Study of the Thylakoid Membrane of a Cyanobacterium Anabaena PCC7120 (2007)
  • Author(s): S. Kumazaki, M. Hasegawa, M. Ghoneim, Y. Shimizu, K. Okamoto, M. Nishiyama, H. Oh-oka, M. Terazima
  • Journal Title: Journal of Microscopy Vol.228 Pages: 240-254
  • Peer Reviewed
• [Journal Article] A Line-Scanning Semiconfocal Multiphoton Fluorescence Microscope with a Simultaneous Broadband Spectral Acquisition and its Application to the Study of the Thylakoid Membrane of a Cyanobacterium Anabaena PCC7120 (2007)
  • Author(s): S. Kumazaki*, M. Hasegawa, M. Ghoneim, Y. Shimizu, K. Okamoto, M. Nishiyama, H. Oh-oka, M. Terazima
  • Journal Title: Journal of Microscopy 228 Pages: 240-254
  • Peer Reviewed
• [Presentation] 顕微蛍光スペクトル測定で捉えたヘテロシスト形成過程のチラコイド膜変化 (2012)
  • Author(s): 熊崎茂一
  • Organizer: 日本植物生理学会年会
  • Place of Presentation: 京都産業大学
  • Year and Date: 2012-03-16
• [Presentation] 顕微蛍光スペクトル測定で捉えたヘテロシスト形成過程のチラコイド膜変化 (2012)
  • Author(s): 明里将志, 長谷川慎, 寺嶋正秀, 熊崎茂一
  • Organizer: 日本植物生理学会
  • Place of Presentation: 京都産業大学
  • Year and Date: 2012-03-16
• [Presentation] 近赤外レーザー励起顕微蛍光スペクトルと顕微吸収スペクトルによる酸素発生型光合成膜の研究 (2012)
  • Author(s): 熊崎茂一
  • Organizer: 私立大学戦略的研究基盤形成支援事業「光合成初期過程器官の時間・空間高分解分光」第1回公開シンポジウム
  • Place of Presentation: 立命館大学(招待講演)
  • Year and Date: 2012-03-13
• [Presentation] アンチストークス蛍光の特性 : 異種葉緑体識別能力と人工的クロロフィル会合体検出 (2012)
  • Author(s): 熊崎茂一
  • Organizer: 分子科学討論会
  • Place of Presentation: 札幌コンベンションセンター
• [Presentation] アンチストークス蛍光の特性:異種葉緑体識別能力と人工的クロロフィル会合体検出 (2011)
  • Author(s): 長谷川慎, 吉田隆彦, 薮田光教, 寺嶋正秀, 熊崎茂一
  • Organizer: 分子科学討論会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2011-09-21
• [Presentation] Anti-Stokes Fluorescence Spectra of Chloroplasts in Green Alga and Plant at Room Temperature as Characterized by Near-Infrared Laser-Based Fluorescence Spectral Microscopy (2011)
  • Author(s): S.Kumazaki, et al
  • Organizer: 5th Asia Oceania Conference on Photobiology
  • Place of Presentation: 奈良県立公会堂
  • Year and Date: 2011-07-31
• [Presentation] 葉緑体個々の構造と機能を同時に見る (2010)
  • Author(s): 熊崎茂一
  • Organizer: 京都植物バイテク懇話会主催、第16回植物バイテクシンポジウム「植物機能を観る、見る、視る、診る、みる」
  • Place of Presentation: 京都府職員研修・研究支援センター(招待講演)
  • Year and Date: 2010-06-17
• [Presentation] Simultaneous Sensing of Photosynthetic Activity and Thylakoid Morphology Realized by Fluorescence and Absorption Spectral Microscopy (2010)
  • Author(s): S. Kumazaki
  • Organizer: The 70th Okazaki International Conference on Molecular mechanism of photosynthetic energy conversion: the present research and future prospects
  • Place of Presentation: Okazaki, Aichi, Japan
• [Presentation] Simultaneous Sellsing of Photosynthetic Activity and Thylakoid Morphology Realized by Fluorescence and Absorption Spectral Microscopy (2010)
  • Author(s): Shigeichi Kumazaki
  • Organizer: The 70th Okazaki International Conference on Molecular mechanism of photosynthetic energy conversion : the present research and future prospects
  • Place of Presentation: Okazaki, Aichi, Japan(招待講演)
• [Presentation] Simultaneous Sensing of Photosynthetic Activity and Thylakoid Morphology Realized by Fluorescence and Absorption Spectral Microscopy (2010)
  • Author(s): Shigeichi Kumazaki
  • Organizer: The 70th Okazaki International Conference on Molecular mechanism of photosynthetic energy conversion : the present research and future prospects
  • Place of Presentation: Okazaki, Aichi, Japan(招待講演)
• [Presentation] 葉緑体内部構造を見るための2光子励起蛍光スペクトル顕微鏡の諸様式 (2009)
  • Author(s): 熊崎茂一
  • Organizer: 日本分光学会生細胞分光部会平成21年度 生細胞分光部会シンポジウム
  • Place of Presentation: 首都大学
  • Year and Date: 2009-12-04
• [Presentation] 葉緑体内部構造を見るための2光子励起蛍光スペクトル顕微鏡の諸様式 (2009)
  • Author(s): 熊崎茂一
  • Organizer: 日本分光学会、生細胞分光部会シンポジウム
  • Place of Presentation: 首都大学東京、八王子、東京
  • Year and Date: 2009-12-04
• [Presentation] 顕微蛍光スペクトルで解析する葉緑体とシアノバクテリアのチラコイド膜自家蛍光 (2008)
  • Author(s): 熊崎茂一
  • Organizer: シンポジウム「見る生物学3」
  • Place of Presentation: 奈良先端科学技術大学院大学、奈良県生駒市
  • Year and Date: 2008-11-14
• [Presentation] Simultaneous broadband fluorescence spectral imaging of a cyanobacterium and chloroplasts facilitated by line-scanning of multiphoton excitation pulses (2008)
  • Author(s): S. Kumazaki
  • Organizer: Focus on Microscopy
  • Place of Presentation: Awaji Yumebutai、Hyogo, Japan
  • Year and Date: 2008-04-15
• [Presentation] 顕微蛍光スペクトルで解析する葉緑体とシアノバクテリアのチラコイド膜自家蛍光 (2008)
  • Author(s): 熊崎茂一
  • Organizer: 奈良先端科学技術大学院大学主催 シンポジウム「見る生物学3」
• [Presentation] 酸素発生型光合成膜微細構造変化を直接捉える顕微分光法 (2008)
  • Author(s): 熊崎茂一
  • Organizer: 分子科学研究書研究会「分子の視点から見る光合成」セミナー(招待講演)分子科学研究所
  • Place of Presentation: 愛知県岡崎市
• [Presentation] Simultaneous broadband fluorescence spectral imaging of a cyanobacterium and chloroplasts facilitated by line-scanning of multiphoton excitation pulses (2008)
  • Author(s): Shigeichi Kumazaki
  • Organizer: Focus on Microscopy 2008
  • Place of Presentation: 淡路島夢舞台
• [Presentation] Micro-regulation of oxygenic photosynthesis unveiled by fluorescence microspectroscopy (2007)
  • Author(s): S. Kumazaki
  • Organizer: The 67th Okazaki Conference, at Okazaki Conference Center, Molecular Science and Chemical Biology of Biomolecular Function
• [Presentation] Micro-regulation of oxygenic photosynthesis unveiled by fluorescence microspectroscopy (2007)
  • Author(s): S. Kumazaki
  • Organizer: The 67th Okazaki Conference "Molecular Science and Chemical Biology of Biomolecular Function"
  • Place of Presentation: Okazaki, Aichi, Japan
• [Presentation] A Line-Scanning Multiphoton Fluorescence Spectromicroscope Applied to the Study of the Thylakoid Membrane in Cyanobacteria and Chloroplasts (2007)
  • Author(s): Shigeichi Kumazaki
  • Organizer: 14th International Congress of Photosynthesis
  • Place of Presentation: Glasgow, Scotland
• [Presentation] A Line-Scanning Semicoufocal Multiphoton Fluorescence Microscope with a Simultaneous Broadband Spectral Acquisition and its Application to the Study of the Thylakoid Membrane of a Cyanobacterium (2007)
  • Author(s): Shigeichi Kumazaki
  • Organizer: state transition
  • Place of Presentation: London, United Kingdom
• [Presentation] Micro-regulation of oxygenic photosynthesis unveiled by fluorescence microspectroscopy (2007)
  • Author(s): Shigeichi Kumazaki
  • Organizer: The The 67th Okazaki Conference, Molecular Science and Chemical Biology of Biomolecular Function
  • Place of Presentation: Okazaki, Japan
• [Book] Three-Dimensional High-resolution Microspectroscopic Study of Environment-Sensitive Photosynthetic Membranes”(Ed. by H.Fukumura, M.Irie, Y.Iwasawa, H.Masuhara and K.Uosaki)in Molecular Nano dynamics, Volume 2 Chapter 30 (2009)
  • Author(s): Shigeichi Kumazaki, Makotoh Hasegawa, Mohammad Ghoneim, Takahiko Yoshida, Masahide Terazima, Takashi Shiina, Isamu Ikegam
  • Publisher: Wiley-VCH
• [Book] 「葉緑体の内部をレーザーで探る」(章著)「反応すれば形が変わるナノの世界～細胞から結晶まで～」 (2009)
  • Author(s): 熊崎茂一
  • Publisher: クバプロ
• [Book] Molecular Nano Dynamics Vol. 2(H. Fukumura, M. Irie, Y. Iwasawa, H. Masuhara and K. Uosaki eds, Three-Dimensional High-resolution Microspectroscopic Study of Environment-Sensitive Photosynthetic Membranes, Chap. 30) (2009)
  • Author(s): S. Kumazaki, M. Hasegawa, M. Ghoneim, T. Yoshida, M. Terazima, T. Shiina, I. Ikegami
  • Publisher: Wiley-VCH
• [Book] 反応すれば形が変わるナノの世界～細胞から結晶まで～(葉緑体の内部をレーザーで探る, ISBN 978-4-87805-099-2) (2009)
  • Author(s): 熊崎茂一
  • Publisher: クバプロ
• [Book] Molecular Nano dynamics, Volume 1, Chapter 16, (Ed. by H. Fukumura, M. Irie, Y. Iwasawa, H. Masuhara and K. Uosaki) (2009)
  • Author(s): S. Kumazaki, M. Hasegawa, M. Ghoneim Yoshida, M. Terazima, T. Shiina, I. Ikegami
  • Total Pages: 20
  • Publisher: Wiley-VCH
• [Book] Photosynthesis. Energy from the Sun, (Ed. by Allen JF, Gantt E, Golbeck JH, and Osmond B.), Volume I, section 2, 93-96 (2009)
  • Author(s): S. Itoh, T. Uzumaki, S. Takaichi, M. Iwaki, S. Kumazaki, K. Itch, and H. Mino
  • Total Pages: 4
  • Publisher: Springer, Dordrecht, The Netherlands
• [Book] Photosynthesis. Energy from the Sun, (Ed. by Allen JF, Gantt E, Golbeck JH, and Osmond B.), Volume I, section 7. 787-790 (2009)
  • Author(s): S. Kumazaki, M. Hasegawa, T. Yoshida, T. Taniguchi, T. Shiina and I. Ikegami
  • Total Pages: 4
  • Publisher: Springer, Dordrecht, The Netherlands
• [Book] A Line-Scanning Multiphoton Fluorescence Spectromicroscope Applied to the Study of the Thylakoid Membrane in Chloroplasts”, Photosynthesis. Energy from the Sun, (Ed. by Allen JF, Gantt E, Golbeck JH, and Osmond B.), Volume I, section 7 (2008)
  • Author(s): Shigeichi. Kumazaki, Makotoh. Hasegawa, Takahiko Yoshida, Tarou Taniguchi, Takashi Shiina and Isamu Ikegami
  • Publisher: Springer, Dordrecht, The Netherlands
• [Book] Energy from the Sun (2008)
  • Author(s): S. Kumazaki*, M. Hasegawa, T. Yoshida, T. Taniguchi, T. Shiina and I. Ikegami
  • Total Pages: 4
  • Publisher: Springer
• [Book] Molecular Nono Dynamics (2008)
  • Author(s): S. Kumazaki*, M. Hasegawa, M. Ghoneim, T. Yoshida, M. Terazima, T. Shiina, I. Ikegami
  • Total Pages: 20
  • Publisher: Wiley-VCH
• [Remarks]
  • URL: http://kuchem.kyoto-u.ac.jp/hikari/kumazaki/microscopic_main.htm