Analyses of functions and interactions of biomolecules by microand nano-devices

• Project/Area Number: 17201031
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Microdevices/Nanodevices
• Research Institution: The University of Tokyo
• Principal Investigator: FUNATSU Takashi The University of Tokyo, 大学院・薬学系研究科, 教授 (00190124)
• Co-Investigator(Kenkyū-buntansha): SANTA Tomofumi 東京大学, 大学院・薬学系研究科, 准教授 (30187306) ; TSUNODA Makoto 東京大学, 大学院・薬学系研究科, 講師 (10323453) ; UENO Taro 東京大学, 大学院・薬学系研究科, 講師 (80376590)
• Project Period (FY): 2005 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥49,790,000 (Direct Cost: ¥38,300,000、Indirect Cost: ¥11,490,000); Fiscal Year 2008: ¥12,480,000 (Direct Cost: ¥9,600,000、Indirect Cost: ¥2,880,000); Fiscal Year 2007: ¥13,780,000 (Direct Cost: ¥10,600,000、Indirect Cost: ¥3,180,000); Fiscal Year 2006: ¥10,920,000 (Direct Cost: ¥8,400,000、Indirect Cost: ¥2,520,000); Fiscal Year 2005: ¥12,610,000 (Direct Cost: ¥9,700,000、Indirect Cost: ¥2,910,000)
• Keywords: 1分子計測 / 生物物理 / ナノバイオ / マイクロ・ナノデバイス / μTAS / セルソーター

Research Abstract

微小流路をチップ上に作製し、蛍光標識した生体試料を蛍光顕微鏡にて高感度に検出し、温度感受性ハイドロゲルをレーザー局所加熱にて制御し、生体試料溶液を高精度に流し分ける技術の開発を行った。その結果、1個の量子ドットに結合した生体分子を高感度に検出し、3msの時間分解能で分離することに成功した。また、石英ガラス基板に金属薄膜を約100nm蒸着し、これに100nmの開口を開けてエバネッセント場を発生させた。このナノ開口を用いてシャペロニンGroELとGroESの結合と解離を観察した。

Research Products

• [Journal Article] On-chip mirofluidic sorting with fluorescence spectrum detection and multiway separation (2009)
  • Author(s): H. Sugino, K. Ozaki, Y. Shirasaki, T. Arakawa, S. Shoji, and T. Funatsu
  • Journal Title: Lab Chip 9 Pages: 1254-60
  • Peer Reviewed
• [Journal Article] Infrared laser-mediated gene induction in targeted single cells in vivo (2009)
  • Author(s): Yasuhiro Kamei, Takashi Funatsu et al.
  • Journal Title: Nature Methods 6 Pages: 79-81
  • Peer Reviewed
• [Journal Article] Hydrophilic fluorescent nanogel thermometer for intracellular thermometry (2009)
  • Author(s): Chie Gota, Takashi Funatsu et al.
  • Journal Title: Journal of American Chemical Society 131 Pages: 2766-2767
• [Journal Article] Single molecule tracking of quantum dot-labeled mRNAs in a cell nucleus (2009)
  • Author(s): Yo Ishihama, Takashi Funatsu
  • Journal Title: Biochemical and Biophysical Research Communications 381 Pages: 33-38
  • Peer Reviewed
• [Journal Article] Real-Time Imaging of Single-Molecule Fluorescence with a Zero-Mode Waveguide for the Analysis of Protein-Protein Interaction (2008)
  • Author(s): T. Miyake, T. Tanii, H. Sonobe, R. Akahori, N. Shimamoto, T. Ueno, T. Funatsu, and I Ohdomari
  • Journal Title: Anal. Chem 80 Pages: 6018-6022
  • Peer Reviewed
• [Journal Article] Real-Time Imaging of Single-Molecule Fluorescence with a Zero-Mode Waveguid for the Analysis of Protein-Protein Interaction (2008)
  • Author(s): Takeo Miyake, Takashi Funatsu et al.
  • Journal Title: Analytical Chemistry 80 Pages: 6018-6022
  • Peer Reviewed
• [Journal Article] Visualization of the molecular dynamics of LPS on the plasma membrane of murine macrophages by TIRFM (2008)
  • Author(s): Samla Shawkat, Takashi Funatsu et al.
  • Journal Title: Journal of Biological Chemistry 283 Pages: 22962-22971
  • Peer Reviewed
• [Journal Article] Football- and bullet-shaped GroEL-GroES complexes coexist during the reaction cycle (2008)
  • Author(s): Tomiya Sameshima, Takashi Funatsu et al.
  • Journal Title: Journal of Biological Chemistry 283 Pages: 23765-23773
  • Peer Reviewed
• [Journal Article] Effect of the C-terminal truncation on the functional cycle of chaperon in GroEL: Implication that the C-terminal region facilitates the transition from the folding-arrested to the folding-competent state (2008)
  • Author(s): Mihoko Suzuki, Takashi Funatsu et al.
  • Journal Title: Journal of Biological Chemistry 283 Pages: 23931-23939
  • Peer Reviewed
• [Journal Article] Kinetic analysis of conformational changes of GroEL based on the fluo rescence of tyrosine 506 (2008)
  • Author(s): Kazuhiko Hosono, Takashi Funatsu et al.
  • Journal Title: The Protein Journal 27 Pages: 461-468
  • Peer Reviewed
• [Journal Article] Rapid multi-reagents exchange TIRFM microfluidic system for a single biomolecular imaging. Sensor Actuat (2007)
  • Author(s): T. Arakawa, T. Sameshima, Y. Sato, T. Ueno, Y. Shirasaki, T. Funatsu, and S. Shoji
  • Journal Title: B-Chem 128 Pages: 218-225
  • Peer Reviewed
• [Journal Article] On-chip Cell Sorting System Using Thermoreversible Gelation Polymer (2007)
  • Author(s): Y. Shirasaki, H. Sugino, M. Tatsuoka, J. Mizuno, S. Shoji, and T. Funatsu
  • Journal Title: IEEE J. Sel. Top. Quant. Electron 13 Pages: 223-227
  • Peer Reviewed
• [Journal Article] Three-dimensional sheath flow sorting microsystem using thermosensitive hydrogel (2007)
  • Author(s): T. Arakawa, Y. Shirasaki, T. Aoki, T. Funatsu, and S. Shoji
  • Journal Title: Sensor Actuat. A-Phys 135 Pages: 99-105
  • Peer Reviewed
• [Journal Article] Selectivity improvement in protein nanopatterning with a hydroxy-terminated self-assembled monolayer template (2007)
  • Author(s): Takeo Miyake, Takashi Funatsu, et. al.
  • Journal Title: Nanotechnology 18
  • Peer Reviewed
• [Journal Article] Rapid multi-reagents exchange TIRFM microfluidic system for a singlebiomolecular imaging (2007)
  • Author(s): Takahiro Arakawa, Takashi Funatsu, et. al.
  • Journal Title: Sensors and Actuators B:Chemical 128 Pages: 218-225
  • Peer Reviewed
• [Journal Article] High-speed particles and biomolecules sorting microsystem using thermosensitive hydrogel (2006)
  • Author(s): T. Arakawa, Y. Shirasaki, T. Izumi, T. Aoki, H. Sugino, T. Funatsu, and S. Shoji
  • Journal Title: Meas. Sci. Techn 17 Pages: 3141-3146
  • Peer Reviewed
• [Journal Article] On-chip cell sorting system using laser-induced heating of a thermo-reversible gelation polymer to control flow (2006)
  • Author(s): Y. Shirasaki, J. Tanaka, H. Makazu, K. Tashiro, S. Shoji, S. Tsukita, and T. Funatsu
  • Journal Title: Anal. Chem 78 Pages: 695-701
  • Peer Reviewed
• [Journal Article] Catecholamine analysis with microcolumn LC-peroxyoxalate chemiluminescence reaction detection (2006)
  • Author(s): Makoto Tsunoda, Takashi Funatsu et al.
  • Journal Title: Clin. Chim. Acta 366 Pages: 168-173
• [Journal Article] A further study on the combined use of internal standard and isotope-labeled derivatization reagent for expansion of linear dynamic ranges in liquid chromatography-electrospray mass spectrometry (2006)
  • Author(s): Yuhki Tsukamoto, Takashi Funatsu et al.
  • Journal Title: Biomed. Chromatogr. 20 Pages: 1049-1055
• [Journal Article] Suppression of thiol exchange reaction in the determination of reduced-form thiols by high-performance liquid chromatography with fluorescence detection after derivatization with fluorogenic benzofurazan reagent, 7-fluoro-2, 1,3-benzoxadiazole-4-sulfonate and 4-aminosulfonyl-7-fluoro-2,1,3- benzoxadiazole (2006)
  • Author(s): Tomofumi Santa, Takashi Funatsu et al.
  • Journal Title: Biomed Chromatogr. 20 Pages: 656-661
• [Journal Article] Localization of Prefoldin Interaction Sites in the Hyperthermophilic Group II Chaperonin and Correlations between Binding Rate and Protein Transfer Rate (2006)
  • Author(s): Tamotsu Zako, Takashi Funatsu et al.
  • Journal Title: J. Mol. Biol. 364 Pages: 110-120
• [Journal Article] High-speed particles and biomolecules sorting microsystem using thermosensitive hydrogel (2006)
  • Author(s): Takahiro Arakawa, Takashi Funatsu et al.
  • Journal Title: Meas. Sci. Techn. 17 Pages: 3141-3146
• [Journal Article] Size distribution of linear and helical polymers in actin solution analyzed by photon counting histogram. (2006)
  • Author(s): Naofumi Terada, Takashi Funatsu et al.
  • Journal Title: Biophys. J. 92 Pages: 2162-2171
• [Journal Article] On-chip cell sorting system using laser-induced heating of a thermo-reversible gelation polymer to control flow (2006)
  • Author(s): Yoshitaka Shirasaki, Takashi Funatsu et al.
  • Journal Title: Anal.Chem 78 Pages: 695-701
• [Journal Article] Synthesis of the isotope-labeled derivatization reagent for carboxylic acids, 7-(N, N-dimethylaminosulfonyl)-4-(aminoethyl)piperazino-2,1,3- benzoxadiazole (d_6) (DBD-PZ-NH_2 (D)), and its application to the quantification and the determination of relative amount of fatty acids in rat plasma samples by high-performance liquid chromatography/mass spectrometry (2006)
  • Author(s): Yuki Tsukamoto, Takashi Funatsu et al.
  • Journal Title: Biomedical Chromatography
• [Journal Article] Nucleocytoplasmic transport of fluorescent mRNA in living mammalian cells : Nuclear mRNA export is coupled to ongoing gene transcription (2006)
  • Author(s): Kazuaki Tokunaga, Takashi Funatsu et al.
  • Journal Title: Genes to Cells 11 Pages: 305-317
• [Journal Article] Rapid assay for catechol-O- methyltransferase activity by high-performance liquid chromatography-fluorescence detection (2005)
  • Author(s): Yoko Hirano, Takashi Funatsu et al.
  • Journal Title: J.Chromatogr.B 819 Pages: 41-46
  • NAID: 10016981128
• [Journal Article] Synthesis of benzofurazan derivatization regents for carboxylic and acids and its application to analysis of fatty acids in rat plasma by high-performance liquid chromatography/electrospray ionization mass spectrometry. (2005)
  • Author(s): Yuki Tsukamoto, Takashi Funatsu et al.
  • Journal Title: Biomed.Chromatogy 19 Pages: 802-808
• [Journal Article] Hybridization of deoxyribonucleic acid immobilization of green fluorescent protein on nanostructured porganosilane templates (2005)
  • Author(s): Takashi Tanii, Takashi Funatsu et al.
  • Journal Title: Jpn.J.App.Phys. 44 Pages: 5851-5855
• [Journal Article] Nanoscale patterning of protein using electron beam lithography of organosilane self-assembled monolayers (2005)
  • Author(s): Guo-Jun Zhang, Takashi Funatsu et al.
  • Journal Title: Small 1 Pages: 833-837
• [Journal Article] Facilitated release of substrate protein from prefoldin by chaperonin (2005)
  • Author(s): Tamotsu Zako, Takashi Funatsu et al.
  • Journal Title: FEBS Lett. 579 Pages: 3718-3724
• [Journal Article] Determination of methylated arginines by column-switching high-performance liquid chromatography-fluorescence detection (2005)
  • Author(s): Makoto Tsunoda, Takashi Funatsu et al.
  • Journal Title: Analyst 130 Pages: 1410-1413
• [Presentation] ナノマイクロデバイスによる生体分子の分離・回収と1分子機能解析 (2008)
  • Author(s): 船津高志, ら
  • Organizer: 第31回日本分子生物学会年会第81回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド、神戸市、兵庫県
  • Year and Date: 2008-12-09
• [Presentation] The long pillar array column with the low-dispersion curve for liquid chromatography (2008)
  • Author(s): Akira Saeki, Takashi Funatsu et al.
  • Organizer: 第18回化学とマイクロ・ナノシステム研究会及びInternational Symposium on Microchemistry and Microsystems 2008合同開催
  • Place of Presentation: 京都大学 桂k
  • Year and Date: 2008-12-07
• [Presentation] PDMSチップのワンステップ3次元鋳造加工と温度感受性ハイドロゲルによる哺乳類細胞の分類 (2008)
  • Author(s): 白崎善隆、船津高志, ら
  • Organizer: 第46回日本生物物理学会年会
  • Place of Presentation: 福岡国際会議場、福岡市、福岡県
  • Year and Date: 2008-12-03
• [Presentation] ナノ開口基板上でのタンパク質翻訳中における後温度のtRNAの1分子ダイナミクス (2008)
  • Author(s): 上村想太郎、船津高志, ら
  • Organizer: 第46回日本生物物理学会年会
  • Place of Presentation: 福岡国際会議場、福岡市、福岡県
  • Year and Date: 2008-12-03
• [Presentation] Cell rupture microfluidic deviceusing nano needle allay for damage-free extraction of organelles (2008)
  • Author(s): Takahiro Arakawa, Takashi Funatsu et al.
  • Organizer: The 12^<yh> International Conference on Miniaturize d systems for Chemistry and Life Sciences 20081012
  • Place of Presentation: San Diego Sheraton Hotel & Marina, Sandiego, California, USA
  • Year and Date: 2008-10-12
• [Presentation] Dynamic imaging of single biomolecular interaction using flow control and TIRFM (2008)
  • Author(s): Takahiro Arakawa, Takashi Funatsu et al.
  • Organizer: The 12^<yh> International Conference on Miniaturize d systems for Chemistry and Life Sciences
  • Place of Presentation: San Diego Sheraton Hotel & Marina, Sandiego, California, USA
  • Year and Date: 2008-10-12
• [Presentation] High performance multiple biomolecules sorting system (2008)
  • Author(s): Kazuto Ozaki, Takashi Funatsu et al.
  • Organizer: The 12^<yh> International Conference on Miniaturize d systems for Chemistry and Life Sciences
  • Place of Presentation: San Diego Sheraton Hotel & Marina, Sandiego, California, USA
  • Year and Date: 2008-10-12
• [Presentation] High performance parallel bioparticle sorter with 3-dimensional PDMS chip (2008)
  • Author(s): Hirokazu Sugino, Takashi Funatsu et al.
  • Organizer: The 12^<th> International Conference on Miniaturize d systems for Chemistry and Life Sciences
  • Place of Presentation: San Diego Sheraton Hotel & Marina, Sandiego, California, USA
  • Year and Date: 2008-10-12
• [Presentation] 生体分子の機能を1分子蛍光イメージングで解き明かす (2008)
  • Author(s): 船津高志
  • Organizer: 日本化学会第2回関東支部大会
  • Place of Presentation: 群馬大学桐生キャンパス、桐生市、群馬県
  • Year and Date: 2008-09-18
• [Presentation] マイクロバルブ流体システムを用いた生体分子-分子イメージ技術の開発 (2008)
  • Author(s): 荒川貴博、船津孝, ら
  • Organizer: 第17回化学とマイクロ・ナノシステム研究会
  • Place of Presentation: 九州大学馬出キャンパス、福岡市、福岡県
  • Year and Date: 2008-05-20
• [Presentation] High performance parallel bioparticle sorter with 3-dimensional PDMS chip (2008)
  • Author(s): H. Sugino, Y. Nara, Y. Shirasaki, T. Arakawa, S. Shoji, and T. Funatsu
  • Organizer: The 12th Int. Conf. on Micro Total Analysis Systems : μTAS2008
  • Place of Presentation: Sandiego, California, USA
• [Presentation] Single molecule imaging of chaperonin functions using zero-mode waveguides (2008)
  • Author(s): Taro Ueno, Takashi Funatsu, et. al.
  • Organizer: Meeting of the Biophysical Society 52nd Annual Meeting and 16th International Biophysics Congress
  • Place of Presentation: Long Beach California,USA
• [Presentation] ADP regulates interaction between Escherichia coli chaperonen GroEL and its cofactor GroES (2008)
  • Author(s): Tomoya Sameshima, Takeshi Funatsu, et.al.
  • Organizer: Meeting of the Biophysical Society 52nd Annual Meeting and 16th International Biophysics Congress
  • Place of Presentation: Long Beach California,USA
• [Presentation] Parallel bioparticle sorting with TGP solution in 3-dimensional microflow system (2007)
  • Author(s): H. Sugino, Y. Nara, Y. Shirasaki, T. Arakawa, S. Shoji and T. Funatsu
  • Organizer: 11th Int. Conf. on Micro Total Analysis Systems : μTAS2007
  • Place of Presentation: Paris, France
• [Presentation] ピラーアレイを用いたオンチップLCの開発 (2007)
  • Author(s): 青山 千顕, 船津 高志, ら
  • Organizer: 日本分析化学会第56年会
  • Place of Presentation: 徳島大学
• [Presentation] An on-chip biomolecule sorting system using laser-induced heating of a thermo-reversible gelation polymer to control flow (2007)
  • Author(s): Takashi Funatsu
  • Organizer: 9th International Symposium on Polymers for advanced Technologie
  • Place of Presentation: Shanghai,China
• [Presentation] Single-molecule analyses of function of biomolecules by nano-and micro-devices (2007)
  • Author(s): Takashi Funatsu
  • Organizer: 第30回日本分子生物学会年会、第80回日本生化学会大会 合同大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] Binding characteristics of thrombopoietin and c-Mpl in Xenopus laevis (2007)
  • Author(s): Yuka Kato, Takashi Funatsu, et. al.
  • Organizer: 第30回日本分子生物学会年会、第80回日本生化学会大会 合同大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] 古細菌シャペロニンの1分子構造変化計測 (2007)
  • Author(s): 飯塚 怜, 船津 高志, ら
  • Organizer: 第30回日本分子生物学会年会、第80回日本生化学会大会 合同大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] ADPがGroELのtransリングへのATPの結合を阻害することにより2個目のGroESが結合できなくなる (2007)
  • Author(s): 鮫島 知哉, 船津 高志, ら
  • Organizer: 第30回日本分子生物学会年会、第80回日本生化学会大会 合同大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] Real time imaging of translocation of fluorescent puromycin-labeled thrombopoietin in Mpl-expressing living cells by fluorescence microscopy (2007)
  • Author(s): Akihiko Sakamoto, Takashi Funatsu, et. al.
  • Organizer: 第30回日本分子生物学会年会、第80回日本生化学会大会 合同大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] GroELのC末端欠損変異はATP加水分解反応を遅くする (2007)
  • Author(s): 鈴木 深保子, 船津 高志, ら
  • Organizer: 日本生物物理学会第45回年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] ADPが対称型1:2GroEL-GroES複合体の形成を阻害する (2007)
  • Author(s): 鮫島 知哉, 船津 高志, ら
  • Organizer: 日本生物物理学会第45回年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] 古細菌シャペロニンの1分子構造変化計測 (2007)
  • Author(s): 飯塚 怜, 船津 高志, ら
  • Organizer: 日本生物物理学会第45回年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] マイクロ流体チップを用いたオルガネラソーティング (2007)
  • Author(s): 杉野 弘和, 船津 高志, ら
  • Organizer: 日本生物物理学会第45回年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] ナノ開口基板を用いたシャペロニンの1分子機能解析 (2007)
  • Author(s): 上野 太郎, 船津 高志, ら
  • Organizer: 日本生物物理学会第45回年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] Accurate and high speed particles and biomolecules sorting microsystem using 3-dimensional sheath flow (2006)
  • Author(s): T. Arakawa, T., T. Aoki, Y. Shirasaki, T. Funatsu, S. Shoji
  • Organizer: 10th Int. Conf. on Micro Total Analysis Systems
  • Place of Presentation: Tokyo, Japan, November
• [Presentation] Analyses of Functions and Interactions of Protein Molecules by Single Fluorescent Molecular Imaging (2006)
  • Author(s): T. Funatsu
  • Organizer: International Conference on Optical MEMS and Their Applications, Big Sky, Montana
  • Place of Presentation: USA, August 21-25
• [Presentation] Rapid Flow Switching with Thermo-Reversible Hydrogel in Microfluidic System (2006)
  • Author(s): Y. Shirasaki, S. Shoji, T. Funatsu
  • Organizer: Biophysical Society Annual Meeting 2006, Salt Palace Convention Center, Salt Lake City,
  • Place of Presentation: Salt Lake City, Utah, USA
• [Presentation] Multi particles and biomolecules sorting systemusing thermoreversible gelation controlled by DMD (2005)
  • Author(s): S. Watabe, M. Tatsuoka, T. Shimomae, Y. Shirasaki, J. Mizuno, T. Funatsu, and S. Shoji
  • Organizer: TRANSDUCERS '05
  • Place of Presentation: Seoul