レーザーを駆使した単一細胞内反応場の構築と計測

2006 Fiscal Year Annual Research Report

• Project/Area Number: 16072211
• Research Institution: Osaka University
• Principal Investigator: 増原 宏 大阪大学, 大学院工学研究科, 教授 (60029551)
• Co-Investigator(Kenkyū-buntansha): 朝日 剛 大阪大学, 大学院工学研究科, 助教授 (20243165) ; 今元 泰 奈良先端科学技術大学院大学, 物質創成科学研究科, 助教授 (80263200) ; 開 祐司 京都大学, 再生医科学研究所, 教授 (40144498)
• Keywords: 単一細胞 / 蛍光イメージング / フェムト秒レーザー / アクチンフィラメント / 緑色蛍光蛋白質 / 衝撃波 / 形態変化

Research Abstract

1.フェムト秒レーザー誘起衝撃波による単一細胞へのナノ粒子導入
マウスNIH3T3細胞から20μm離れた位置にフェムト秒パルスを照射すると細胞は変形するが生きている。次に200nmの蛍光性高分子微粒子を添加し同条件下でパルス励起し、細胞内に導入されることを3次元蛍光イメージングで示し、新しい方法論として提案した。
2.フェムト秒レーザー直接照射による単一細胞へのDNAプラスミドの導入と発現
DNAプラスミドのモデルとして蛍光分子をラベルしたデキストランを用い、レーザー照射による導入の条件を明らかにした。また緑色蛍光蛋白プラスミドを導入し、発現させることに成功した。後者の場合、細胞膜にのみ集光するだけではなく、核膜をも照射したときにのみ核外に発現を確認しており、その導入と発現のメカニズムについて調べた。
3.単一金ナノ粒子プラズモン共鳴分光・イメージングと生体細胞への応用
フェムト秒白色光をプローブ光として共焦点顕微システムを用い、マウスNIH3T3細胞の3次元光散乱イメージングと分光を行った。細胞のみの場合、細胞構造を反映した3次元光散乱画像のスペクトルには特徴あるピークは検出されなかった。金ナノ粒子を添加するとプラズモン共鳴バンドが見られ、単一粒子レベルで分布と会合状態を考察できるようになった。これとは別に、プローブである金ナノ粒子のスペクトルに及ぼす形状と吸着色素の影響を明らかにし、その機構を検討した。
4.共同研究の成果としては笹木研(A01班)と単一微粒子分光の研究、佐々木研(A04班)とfsパルス誘起結晶化の研究、宮坂研(A04班)とフェムト秒多光子顕微鏡の開発と応用の研究が既に原著論文として発表印刷されている。

Research Products

• [Journal Article] Dual Effects of the Membrane-anchored MMP Regulator RECK on Chondrogenic Differentiation of ATDC5 Cell (2007)
  • Author(s): Shunya Kondo, Chisa Shukunami, Yoko Morioka, Naoya Matsumoto, Rei Takahashi, Junseo Oh, Tadao Atsumi, Akihiro Umezawa, Akira Kudo, Hitoshi Kitayama, Yuji Hiraki, Makoto Noda
  • Journal Title: J. Cell Sci. (in press)
• [Journal Article] Anthracene Crystallization Induced by Single-shot Femtosecond Laser Irradiation : Experimental Evidence for the Important Role of Bubbles (2007)
  • Author(s): Kazuhiko Nakamura, Yoichiroh Hosokawa, Hiroshi Masuhara
  • Journal Title: Crystal Growth and Design (in press)
• [Journal Article] Polarization and Wavelength Dependent Nonlinear Optical Properties of a Photo-Switchable Organic Crystal (2007)
  • Author(s): Michel Sliwa, Keitaro Nakatani, Tsuyoshi Asahi, Pascal G. Lacroix, Robert B. Pansu, Hiroshi Masuhara
  • Journal Title: Chem. Phys. Let. (in press)
• [Journal Article] In situ Micro-patterning of Proteinous Occlusion Bodies in Water by Femtosecond Laser-Induced Mechanical Fore (2007)
  • Author(s): Yoichiroh Hosokawa, Takahiro Kaji, Chisa Shukunami, Yuji Hiraki, Eiji Kotani, Hajime Mori, Hiroshi Masuhara
  • Journal Title: Biomedical Microdevices (in press)
• [Journal Article] Femtosecond Laser-induced Cleaving of Protein Crystal in Water Solution (2007)
  • Author(s): Masafumi Kashii, Yoichiroh Hosokawa, Hiroshi Kitano, Hiroaki Adachi, Yusuke Mori, Kazufumi Takano, Hiroyoshi Matsumura, Tsuyoshi Inoue, Satoshi Murakami, Kazuomi Sugamoto, Hideki Yoshikawa, Takatomo Sasaki, Hiroshi Masuhara
  • Journal Title: Appl. Surface Sci. (in press)
• [Journal Article] Femtosecond Laser-induced Crystallization of Protein in Gel Medium (2007)
  • Author(s): Kazuhiko Nakamura, Yosuke Sora, Hiroshi Y.Yoshikawa, Yoichiroh Hosokawa, Yusuke Mori, Takatomo Sasaki, Hiroshi Masuhara
  • Journal Title: Appl. Surface Sci. (in press)
• [Journal Article] Realignment Process of Actin Stress Fibers in Single Living Cells Studied by Focused Femtosecond Laser Irradiation (2007)
  • Author(s): Ryohei Yasukuni, Jean-Alexis Spitz, Rachel Meallet-Renault, Takayuki Negishi, Takuji Tada, Yoichiro Hosokawa, Tsuyoshi Asahi, Chisa Shukunami, Yuji Hiraki, Hiroshi Masuhara
  • Journal Title: Appl. Surface Sci. (in press)
• [Journal Article] Spatial Light Modulating and Multi-trapping with a DMD (2007)
  • Author(s): Yuqiang Jiang, Yong Wang, Yoshikata Matsumoto, Yoichiroh Hosokawa, Hiroshi Masuhara
  • Journal Title: Modern Phys. Lett. B (in press)
• [Journal Article] Laser ablation of Individual Gold Nanoparticles in Solution (2007)
  • Author(s): Syoji Ito, Toshiaki Mizuno, Hiroyuki Yoshikawa, Hiroshi Masuhara
  • Journal Title: Jpn. J. Appl. Phys. Vol.46, No. 10 Pages: L241-L243
• [Journal Article] Trapping and Manipulation of a Single Micro-object in Solution with Femtosecond Laser-induced Mechanical Force (2007)
  • Author(s): Yuqiang Jiang, Yoshitaka Matsumoto, Yoichiroh Hosokawa, Hiroshi Masuhara, Isamu Oh
  • Journal Title: Appl. Phys. Lett. Vol. 90. No.6 Pages: 061107-1-061107-3
• [Journal Article] Multipole Resonance Modes in Localized Surface Plasmon of Single Hexagonal/Triangular Gold Nanoplate (2007)
  • Author(s): Takayuki Uwada, Tsuyoshi Asahi, Hiroshi Masuhara, Daisuke Ibano, Manabu Fujishiro, Toshihiro Tominaga
  • Journal Title: Chem. Lett. Vol.36, No.2 Pages: 318-319
• [Journal Article] ndividual Bonding of Biological Micromaterials in Water by UV Laser-Induced Polymerization of a Resin Layer in Water (2006)
  • Author(s): Chie Matsubara, Yoichiroh Hosokawa, Yong Wang, Hiroshi Masuhara
  • Journal Title: Indian J. Radiation Research Vol. 3, No. 2＆3 Pages: 94-98
• [Journal Article] Scleraxis Positively Regulates the Lineage-specific Expression of Tenomodulin, a Marker of Tenocytes (2006)
  • Author(s): Chisa Shukunami, Aki Takimoto, Miwa Oro, Yuji Hiraki
  • Journal Title: Dev. Biol. Vol. 298, No. 1 Pages: 234-247
• [Journal Article] Laser Nano Chemistry (2006)
  • Author(s): Hiroshi Masuhara, Tsuyoshi Asahi, Yoichiroh Hosokawa
  • Journal Title: Pure Appl. Chem. Vol. 78, No. 12 Pages: 2205-2226
• [Journal Article] Chondromodulin-I Maintains Cardiac Valvular Function by Preventing Angiogenesis (2006)
  • Author(s): Masatoyo Yoshioka, Shinsuke Yuasa, Keisuke Matsumura, Takayuki Shiomi, Chisa Shukunami, Yasunori Okada, Makio Mukai, Hankei Shin, Ryohei Yozu, Masataka Sata, Satoshi Ogawa, Yuji Hiraki, Keiichi Fukuda
  • Journal Title: Nature Medicine Vol. 12 No. 10 Pages: 1151-1159
• [Journal Article] Stilbene Analogs in Hula-Twist Photoisomerization (2006)
  • Author(s): Lan-Ying Yang, Miki Harigai, Yasushi Imamoto, Mikio Kataoka, Tong-Ing Ho, Elena Andrioukhina, Olga Federova, Sergei Shevyakov, Robert S. H. Liu
  • Journal Title: Photochem. Photobiol. Sci. Vol. 5, No. 10 Pages: 874-882
• [Journal Article] Organic nonlinear optical DAST crystals for electro-optic measurement and terahertz wave generation (2006)
  • Author(s): Yoshinori Takahashi, Hiroaki Adachi, Tetsuo Taniuchi, Masayuki Takagi, Youichiroh Hosokawa, Shinji Onzuka, Srinivasan Brahadeeswaran, Masashi Yoshimura, Yusuke Mori, Hiroshi Masuhara, Takatomo Sasaki, Hachiro Nakanishi
  • Journal Title: J. Photochem. Photobiol. A : Chem. Vol.183, No.3 Pages: 247-252
• [Journal Article] Higher-order Multiphoton Imaging by Femtosecond Near-Infrared Laser Microscope System (2006)
  • Author(s): Hirohisa Matsuda, Syoji Ito, Yutaka Nagasawa, Tsuyoshi Asahi, Hiroshi Masuhara, Seiya Kobatake, Masahiro Irie, Hiroshi Miyasaka
  • Journal Title: Photochem. Photobiol. A : Chem. Vol. 183, No.3 Pages: 261-266
• [Journal Article] Expression of the PTH/PTHrP Receptor in Chondrogenic Cells During the Repair of Full-Thickness Defects of Articular Cartilage (2006)
  • Author(s): Hiroshi Mizuta, Satoshi Kudo, Eiichi Nakamura, Katsumasa Takagi, Yuji Hiraki
  • Journal Title: Osteoarthritis Cartilage Vol. 14, No. 9 Pages: 944-952
• [Journal Article] Trans-cis Photoisomerization of a Photoactive Yellow Protein Model Chromophore in Crystalline Phase (2006)
  • Author(s): Anwar Usman, Hiroshi Masuhara, Tsuyoshi Asahi
  • Journal Title: J. Phys. Chem. B Vol. 110, No. 41 Pages: 20085-20088
• [Journal Article] A Single CH/□ Weak Hydrogen Bond Governs Stability and the Photocycle of the Photoactive Yellow Protein (2006)
  • Author(s): Miki Harigai, Mikio Kataoka, Yasushi Imamoto
  • Journal Title: Am. Chem. Soc. Vol. 128, No. 33 Pages: 10646-10647
• [Journal Article] Spectroscopy and Photochemistry of Single Organic Nanocrystals Investigated by Using a Far-Eield Optical Microscope Coupled with an AFM System (2006)
  • Author(s): Tsuyoshi Asahi, Victor V.Volkov, Hideki Matsune, Hidetsugu Kawai, Hiroshi Masuhara
  • Journal Title: Proc. of Electron Transfer in Nanomaterials Vol. 2004-22 Pages: 150-161
• [Journal Article] Two-Photon Fluorescence Spectroscopy of Individually Trapped Pseudoisocyanine J-Aggregates in Aqueous Solution (2006)
  • Author(s): Yoshito Tanaka, Hiroyuki Yoshikawa, Hiroshi Masuhara
  • Journal Title: J. Phys. Chem. B Vol. 110, No. 36 Pages: 17906-17911
• [Journal Article] Conformational Changes of PYP Monitored by Diffusion Coefficient : Effect of N-terminal □-Helices (2006)
  • Author(s): Javaid Shahbaz Khan, Yasushi Imamoto, Miki Harigai, Mikio Kataoka, Masahide Terazima
  • Journal Title: Biophys. J. Vol. 90, No. 10 Pages: 3686-3693
• [Journal Article] Enhancement of Biased Diffusion of Dye-Doped Nanoparticles by Simultaneous Irradiation with Resonance and Nonresonance Laser Beams (2006)
  • Author(s): Chie Hosokawa, Hiroyuki Yoshikawa, Hiroshi Masuhara
  • Journal Title: Jpn. J. Appl. Phys. Vol. 45, No. 16 Pages: L453-L456
• [Journal Article] The Crystal Structure of the R52Q Mutant Demonstrates a Role for R52 in Chromophore pKa Regulation in Photoactive Yellow Protei (2006)
  • Author(s): Nobutaka Shimizu, Hiroari Kamikubo, Yoichi Yamazaki, Yasushi Imamoto, Mikio Kataoka
  • Journal Title: Biochemistry Vol. 45, No. 11 Pages: 3542-3547
• [Book] Nano Biophotonics : Science and Technology (第15,16,22,24章) (2007)
  • Author(s): Hiroshi Masuhara, Tsuyoshi Asahi, Yuuji Hiraki, et al.
  • Total Pages: 428
  • Publisher: Elsevier