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Production of novel proteins using the micro system that permits sorting and functional evaluation at single molecular level

Research Project

Project/Area Number 13558088
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Biophysics
Research InstitutionWaseda University

Principal Investigator

FUNATSU Takashi  Waseda University, School of Science and Engineering, Professor, 理工学部, 教授 (00190124)

Co-Investigator(Kenkyū-buntansha) TSUKITA Shoichiro  Kyoto University, Graduate School of Medicine, Professor, 大学院・医学研究科, 教授 (50155347)
SHOJI Shuichi  Waseda University, School of Science and Engineering, Professor, 理工学部, 教授 (00171017)
根本 直人  (株)GEN COM, 研究開発1室, 主任研究員
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2003: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2002: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2001: ¥7,300,000 (Direct Cost: ¥7,300,000)
KeywordsMicro-system / Molecule Sorter / Single-molecule Probe
Research Abstract

We have developed a micro fabricated fluorescence-activated bio-molecular sorter using thermosensitive hydrogel as a microvalve. The sorter was composed of a microfluidic chip with Y-shaped channels with one inlet and two outlets. Biological specimen containing fluorescently labeled cells and thermosensitive polymer was introduced into a Y-shaped microchannel. Thermosensitive polymer changed the state from sol to gel when it was heated. The sol-gel transition was locally induced by site directed infrared laser irradiation in order to plug the one of the microchannel and collect the cells. As an application of this sorter, GFP-expressing E. coll. CD8-expressing lymphocytes labeled with fluorescence antibody, and DNA molecules labeled with TOTO-1 were separated. The sorting time of 120 ms and recovery ratio of 90 % was achieved by this sorter.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (46 results)

All Other

All Publications (46 results)

  • [Publications] Yamaguchi, J. et al.: "Rapid functional analysis of protein-protein interactions by fluorescent C-terminal labeling and single-molecule imaging"FEBS Lett.. 502. 79-83 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Taguchi, H. et al.: "Single-molecule observation of protein-protein interactions in the chaperonin system."Nature Biotechnology. 19. 861-865 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tadakuma, H. et al.: "Imaging of Single Fluorescent Molecules Using Video-rate Confocal Microscopy."Biochem Biophys Res Commun.. 287. 323-327 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tashiro, K., et al.: "A Particles and Biomolecules Sorting Micro Flow System Using Thermal Gelation of Methyl Cellulose Solution."Micro Total Analysis Systems 2001. 471-473 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 多田隈尚史, 船津高志: "生きた細胞の核内mRNAの1分子蛍光イメージング"細胞工学. 20. 672-677 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tashiro, K., et al.: "Micro Flow Switches Using Thermal Gelation of Methyl Cellulose for Biomolecules Handling."11th International Conference on Solid-State Sensors and Actuators, (Transducers'01). 932-935 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Funatau et al.: "Rapid and Sensitive Detection Method of A Bacterium Using GFP Reporter Phage."Microbiol.Immuno.. 6. 365-369 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Fujiwara et al.: "Microscopicanalysis of polymerization dynamics with individual actin filaments."Nature Cell Biol.. 4. 666-673 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shirasaki et al.: "A Novel Biomolecule Sorter Using Thermosensitve Hydrogel in Micro Flow System"Micro Total Analysis System 2002. 925-927 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 船津高志: "1分子蛍光イメージング技術による生命現象の解析"可視化情報学会誌. 22. 18-21 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 座古保, 船津高志: "生体機能を1分子レベルで探る -1分子蛍光イメージング-"分光研究. 51. 3-14 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kanai et al.: "PDMS microfluidics devices with PTFE passivated channels."Micro Total Analysis System 2003. 429-432 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 船津高志: "単一分子計測とバイオフォトニクス"応用物理. 72. 727-730 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 多田隈尚史ら: "mRNA核外輸送の可視化"蛋白質核酸酵素. 48. 421-429 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Zhang, G. et al.: "The immobilization of DNA on microstructured patterns fabricated by maskless lithography."Sensors and Actuators B. 97. 243-248 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Zhang, G. et al.: "Patterning of DNA nanostructures on silicon surface by electron beam lithography of self-assembled monolayer."Chem Comm.. 786-787 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ueno, T. et al.: "GroEL mediates protein folding with a two successive timer mechanism."Mol.Cell. (in press). (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yamaguchi, J., N.Nemoto, T.Sasaki, A.Tokumasu, Y.Mimori-Kiyorue, T.Yagi, T.Funatsu: "Rapid functional analysis of protein-protein interactions by fluorescent C-terminal labeling and single-molecule imaging"FEES Lett. 502. 79-83 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Taguchi, H., T.Ueno, H.Tadakuma, M.Yoshida, T.Funatsu: "Single-molecule observation of protein-protein interactions in the chaperonin system"Nature Biotechnology. 19. 861-865

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tadakuma, H., J.Yamaguchi, Y.Ishihama, T.Funatsu: "Imaging of Single Fluorescent Molecules Using Video-rate Con focal Microscopy"Biochem Biophys Res Commun.. 287. 323-327 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tashiro, K., S.Ikeda, T.Sekiguchi, S.Shoji, H.Makazu, T.Funatsu, S.Tsukita: "A Particles and Biomolecules Sorting Micro Flow System Using Thermal Gelation of Methyl Cellulose Solution"Micro Total Analysis Systems 2001. 471-473 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tashiro, K., S.Ikeda, T.Sekiguchi, H.Sato, S.Shoji, H.Makazu, K.Watanabe, T.Funatsu, S.Tsukita.: "Micro Flow Switches Using Thermal Gelation of Methyl Cellulose for Biomolecules Handling"11th International Conference on Solid-State Sensors and Actuators. Transducers'01. 932-935 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Funatsu, T., T.Taniyama, T.Tajima, H.Tadakuma, H.Namiki: "Rapid and Sensitive Detection Method of A Bacterium Using GFP Reporter Phage"Microbiol. Immune. 46. 365-369 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Fujiwara, I., S.Takahashi, H.Tadakuma, T.Funatsu, S.Ishiwata: "Microscopicanalysis of polymerization dynamics with individual actin filaments"Nature Cell Biol.. 4. 666-673 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shirasaki, Y., T.Funatsu: "A Novel Biomolecule Sorter Using Thermosensitve Hydrogel in Micro Flow System"Micro Total Analysis System 2002. 925-927 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kanai, M., D.Uchida, S.Sugiura, Y.Shirasaki, J.S.Go, H.Nakanishi, T.Funatsu, S.Shoji: "PDMS microfluidics devices with PTFE passivated channels"Micro Total Analysis System 2003. 429-432 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Zhang, G., T.Tanii, T.Zako, T.Funatsu, I.Ohdomari: "The immobilization of DNA on microstructured patterns fabricated by maskless lithography"Sensors and Actuators B. 97. 243-248 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Zhang, G.-J., T.Tanii, T.Funatsu, I.Ohdomari: "Patterning of DNA nanostructures on silicon surface by electron beam lithography of self-assembled monolayer"Chem Comm. 786-787 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ueno, T., H.Taguchi, H.Tadakuma, M.Yoshida, T.Funatsu.: "GroEL mediates protein folding with a two successive timer mechanism"Mol. Cell 2004. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] G.Zhang: "The immobilization of DNA on microstructured patterns fabricated by maskless lithography"Sensors and Actuators B. 97. 243-248 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] G.Zhang: "Patterning of DNA nanostructures on silicon surface by electron beam lithography of self-assembled monolayer."Chem Comm.. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] M.Kanai: "PDMS microfluidics devices with PTFE passivated channels."Micro Total Analysis System 2003. 429-432 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 船津高志: "単一分子計測とバイオフォトニクス"応用物理. 72. 727-730 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] S.Shoji: "Particles and molecules handling in micro channels"Lab-on-a-Chip.Miniaturized Systems for (Bio)Chemical Analysis and Synthesis.. 205-214 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 多田隈尚史: "mRNA核外輪送の可視化"蛋白質核酸酵素. 48. 421-429 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Funatsu, T. et al.: "Rapid and sensitive detection method of a bacterium using GFP reporter phage"Microbiol.Immuno.. 46. 365-369 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Fujiwara, I. et al.: "Microscopic analysis of polymerization dynamics with individual actin filaments"Nature Cell Biol.. 4. 666-673 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shirasaki, Y. et al.: "A novel biomolecule sorter using thermosensitive hydrogel in micro flow system"Micro Total Analysis System 2002. 925-927 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 船津高志: "1分子蛍光イメージング技術による生命現象の解析"可視化情報学会誌. 22. 18-21 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 座古保, 船津高志: "生体機能を1分子レベルで探る"分光研究. 51. 3-14 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yamaguchi, J. et al.: "Rapid functional analysis of protein-protein interactions by fluorescent C-terminal labeling and single-molecule imaging"FEBS Lett. 502. 79-83 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Taguchi, H. et al.: "Single-molecule observation of protein-protein interactions in the chaperonin system"Nature Biotechnology. 19. 861-865 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tadakuma, H. et al.: "Imaging of Single Fluorescent Molecules Using Video-rate Confocal Microscopy"Biochem Biophys Res Commun. 287. 323-327 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tashiro, K., et al.: "A Particles and Biomolecules Sorting Micro Flow System Using Thermal Gelation of Methyl Cellulose Solution"Micro Total Analysis Systems 2001. 471-473 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 多田隈尚史, 船津高志: "生きた細胞の核内mRNAの1分子蛍光イメージング"細胞工学. 20. 672-677 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tashiro, K., et al.: "Micro Flow Switches Using Thermal Gelation of Methyl Cellulose for Biomolecules Handling"11th International Conference on Solid-State Sensors and Actuators,(Transducers'O1). 932-935 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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