Micro-culture device for environmental condition control and quantitative electrochemical analysis

• Project/Area Number: 19750055
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Analytical chemistry
• Research Institution: Tohoku University
• Principal Investigator: SHIKU Hitoshi Tohoku University, 大学院・環境科学研究科, 准教授 (10361164)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,780,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥480,000); Fiscal Year 2008: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2007: ¥1,700,000 (Direct Cost: ¥1,700,000)
• Keywords: チップ分析 / 電気化学 / レポーター遺伝子 / 細胞培養

Research Abstract

我々はこれまでに、ベータ・ガラクトシダーゼ(βGAL)およびアルカリホスファターゼ(AP)をレポーター酵素とし、遺伝子発現のon/offにより制御されるシグナル伝達の電気化学的追跡が可能であることを示した。本課題では、単一細胞レベルでの酵素活性検出を目標とする。これを実現するために、分泌型酵素を利用しレポーター酵素を膜外に分泌させ、大幅な感度の向上を目指す(分泌型APの適用)。電極・ウェルデザインの最適化により酵素反応生成物が効率的に電極に到達するよう検討を加える。

Research Products

• [Journal Article] Development of Electrochemical Reporter Assay Using HeLa Cells Transfected with Vector PLasmids Encoding Various Responsive Elements (2009)
  • Author(s): H. Shiku, M. Takeda, T. Murata, U. Akiba, F. Hamada, T. Matsue
  • Journal Title: Anal. Chim. Acta 640 Pages: 87-92
  • Peer Reviewed
• [Journal Article] Electrochcmical Charactcrization of Enzymatic Activity of Ycast Cells Entrapped in a Poly(dimethylsiloxane)Microwell on the Basis of Limited Diffusion System (2009)
  • Author(s): H. Shiku, S. Goto, S. Jung, K. Nagamine, M. Koide, T. Itayama, T. Yasukawa, T. Matsue
  • Journal Title: Analyst 134 Pages: 182-187
  • Peer Reviewed
• [Journal Article] A microfluidic dual capillary probe to collect messenger RNA from adherent cells and spheroids (2009)
  • Author(s): H. Shiku, T. Yamakawa, Y. Nashimoto, Y. Takahashi, Y. Torisawa, T. Yasukawa, T. Ito-Sasaki, M. Yokoo, H. Abe, H. Kambara, T. Matsue
  • Journal Title: Analytical Biochemistry 385 Pages: 138-142
  • Peer Reviewed
• [Journal Article] Entrapment and Measurement of a Biologically Functionalized Microbead with a Microwell Electrode (2009)
  • Author(s): C. -Y. Chang, T. Murata, Y. Takahashi, H. Shiku, H. -C. Chang, T. Matsue
  • Journal Title: Lab Chip 2009,9 Pages: 1185-1192
  • Peer Reviewed
• [Journal Article] Electrochemical Detection of Epidermal Growth Factor Receptors on a Single Living Cell Surface by Scanning Electrochemical Microscopy (2009)
  • Author(s): . Takahashi, T. Miyamoto, H. Shiku, R. Asano, T. Yasukawa, I. Kumagai, T. Matsue
  • Journal Title: Anal. Chem. 81(7) Pages: 2785-2790
  • Peer Reviewed
• [Journal Article] Electrochemical Characterization of Enzymatic Activity of Yeast Cells Entrapped in a Poly (dimethylsiloxane) Microwell on the Basis of Limited Diffusion System (2009)
  • Author(s): H. Shiku, et.al.
  • Journal Title: Analyst 134 Pages: 182-187
  • Peer Reviewed
• [Journal Article] A microfluidic dual capillary probe to collect messenger RNA from adherent cells and spheroidsAnalytical Biochemistry (2009)
  • Author(s): H. Shiku, et.al.
  • Journal Title: Analytical Biochemistry 385 Pages: 138-142
  • Peer Reviewed
• [Journal Article] Electrochemical gene-function analysis for single cells with addressable electrode/microwell arrays (2009)
  • Author(s): Z. Lin, et.al.
  • Journal Title: Angew. Chem. Mt. Ed. 48 Pages: 2044-2046
  • Peer Reviewed
• [Journal Article] Entrapment and Measurement of a Biologically Functionalized Microbead with a Microwell Electrode (2009)
  • Author(s): C.-Y. Chang, et.al.
  • Journal Title: Lab Chip 9(印刷中)
  • Peer Reviewed
• [Journal Article] Electrochemical Detection of Epidermal Growth Factor Receptors on a Si ngle Living Cell Surface by Scanning Electrochemical Microscopy (2009)
  • Author(s): Y. Takahashi, et.al.
  • Journal Title: Anal. Chem 81 Pages: 2785-2790
  • Peer Reviewed
• [Journal Article] Development of Electrochemical Reporter Assay Using HeLa Cells Transfected with Vector Plasmids Encoding Various Responsive Elements (2009)
  • Author(s): H. Shiku, et.al.
  • Journal Title: Anal. Chim. Acta 640 Pages: 87-92
  • Peer Reviewed
• [Journal Article] Electrophoretic cell manipulation and elettrochemical gene-function analys is based on a yeast two-hybrid system in a microfluidic device (2008)
  • Author(s): T. Yasukawa, et.al.
  • Journal Title: Anal. Chem 80 Pages: 3722-3727
  • Peer Reviewed
• [Journal Article] An addressable microelectrode array for electrochemical detection (2008)
  • Author(s): Z. Lin, et.al.
  • Journal Title: Anal. Chem 80 Pages: 6830-6833
  • Peer Reviewed
• [Journal Article] Microfluidic chip integrated with amperometric detector array for in situ estimating oxygen consumption characteristics of single bovine embryos (2007)
  • Author(s): C. -C. Wu, T. Saito, T. Yasukawa, H. Shiku, H. Abe, H. Hoshi, T. Matsue
  • Journal Title: Sens. Actuat. B 125 Pages: 680-687
  • Peer Reviewed
• [Journal Article] Electrochemical screening of recombinant protein solubility in Escherichia coli using scanning electrochemical microscopy(SECM) (2007)
  • Author(s): K. Nagamine, et. al.
  • Journal Title: Biotechnol. Bioeng. 96 Pages: 1008-1013
  • Peer Reviewed
• [Journal Article] A multicellular spheroid array chip to realize spheroid formation, culture and viability assay (2007)
  • Author(s): Y. Torisawa, et. al.
  • Journal Title: Biomaterials 28 Pages: 559-566
  • Peer Reviewed
• [Journal Article] Microfluidic chip integrated with amperometric detector array for in situ estimating oxygen consumption characteristics of single bovine embryos (2007)
  • Author(s): Y. Nashimoto, et. al.
  • Journal Title: Analyitical Chemistry 79 Pages: 6823-6830
  • Peer Reviewed
• [Presentation] Single-cell analysis based on scanning probe technique and electrochemical microdevice (2009)
  • Author(s): H. Shiku
  • Organizer: 日本化学会第89回年会アジア国際シンポジウム
  • Place of Presentation: 日本大学理工学部舟橋キャンパス(招待講演)
  • Year and Date: 2009-03-29
• [Presentation] 一細胞採取およびPCR解析に基づくデジタル解析 (2009)
  • Author(s): 珠玖 仁
  • Organizer: 第8回日本再生医療学会総会
  • Place of Presentation: 東京国際フォーラム, 東京都千代田区(招待講演)
  • Year and Date: 2009-03-05
• [Presentation] 一細胞採取およびPCR解析に基づくデジタル解析 (2009)
  • Author(s): 珠玖仁
  • Organizer: 第8回日本再生医療学会総会
  • Place of Presentation: 東京国際フォーラム, 東京都(招待講演)
  • Year and Date: 2009-03-05
• [Presentation] Single-cell gene expression analysis based on scanning probe microscopy (2008)
  • Author(s): H. Shiku, Y. Takahashi, T. Murata, T. Matsue
  • Organizer: IUMRS international conference in Asia 2008
  • Place of Presentation: Nagoya Congress Center, Nagoya
  • Year and Date: 2008-12-12
• [Presentation] SECM (scanning electrochemical microscopy) for single cell bioimaging (2008)
  • Author(s): H. Shiku
  • Organizer: PRiME 2008, 214th Meeting of The Electrochemical Society
  • Place of Presentation: Honolulu, Hawaii (USA)
  • Year and Date: 2008-10-16
• [Presentation] Scanning Probe technique and electrochemical microdevice for single cell analysis (2008)
  • Author(s): H. Shiku
  • Organizer: JAIST Nano Technology Symposium 2008
  • Place of Presentation: Ishikawa High-Tech Conference Center, Ishikawa(招待講演)
• [Presentation] Electrochemical Analysis of Yeast Cells Expressing Beta-Galactosidase within a Poly (dimethylsiloxane) Microwell Array (2008)
  • Author(s): H. Shiku, S. Goto, K. Nagamine, T. Yasukawa, T. Itayama, T. Matsue
  • Organizer: 5^<th> Workshop on Scanning electrochemical Microscopy
  • Place of Presentation: Blue Mountain Lake, NY (USA)
• [Presentation] マイクロ・コンタクトプリンティング法におけるタンパク質パターニング効率の電気化学的定量評価 (2008)
  • Author(s): 珠玖 仁,熊谷 絢子,羅 紅群,高橋 康史,安川 智之,山田 弘,末永 智一
  • Organizer: 電気化学会第75回大会
  • Place of Presentation: 山梨大学
• [Presentation] 密閉系マイクロウェルアレイによる酵母の電気化学評価 (2008)
  • Author(s): 珠玖 仁,後藤 俊,チョン ソンボン,長峯 邦明,安川 智之,末永 智一
  • Organizer: 電気化学会第75回大会
  • Place of Presentation: 山梨大学
• [Presentation] Oxygen Consumption of Mammalian Embryos and Oocytes Monitored by Scanning Electrochemical Microscopy (2007)
  • Author(s): H. Shiku, T. Yasukawa, T. Matsue, T. Ito-Sasaki, M. Yokoo, H. Abe
  • Organizer: IEEE Sensors 2007
  • Place of Presentation: Atlanta Georgia USA
• [Presentation] 単一細胞由来mRNA回収プローブと電気化学マイクロシステムの開発(招待講演) (2007)
  • Author(s): 珠玖 仁
  • Organizer: 生体機能関連化学部会若手の会サマースクール
  • Place of Presentation: 八王子セミナーハウス、東京
• [Book] Bioelectrochemstry : fundamentals, experimental techniques, and applications (2008)
  • Author(s): H. Shiku, K. Nagamine, T. Kaya, T. Yasukawa, T. Matsue
  • Publisher: Chapter 7, John Wiley & Sons Ltd., West Sussex, UK. (Eds. P. N. Bartlett)
• [Book] John Wiley & Sons Ltd., West Sussex, UK.(Eds.P.N.Bartlett) (2008)
  • Author(s): H. Shiku, et.al.
  • Publisher: Bioelectrochemistry : fundamentals, experimental techniques, and applications, Chapter 7
• [Book] 「生体分子の局所分析と電気化学イメージング」、(バイオ電気化学の実際-バイオセンサ・バイオ電池の実用展開-、監修 池田篤治 、第13章 (2007)
  • Author(s): 高橋康史, 他
  • Publisher: シーエムシー出版
• [Remarks]
  • URL: http://db.tohoku.ac.jp/whois/detail/acd9a15637a319e79930f9ff86a2b16c.html