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

2008 Fiscal Year Final Research Report

• 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
• 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] 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
• [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] 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)
  • Year and Date: 20081023-25
• [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)
  • Year and Date: 20080824-28
• [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
  • Year and Date: 20080328-30
• [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
  • Year and Date: 20080328-30
• [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] 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
  • Year and Date: 20071028-31
• [Book] Bioelectrochemstry : fundamentals, experimental techniques, and applications (2008)
  • Author(s): H. Shiku, K. Nagamine, T. Kaya, T. Yasukawa, T. Matsue
  • Total Pages: 249-266
  • Publisher: Chapter 7, John Wiley & Sons Ltd., West Sussex, UK. (Eds. P. N. Bartlett)
• [Remarks]
  • URL: http://db.tohoku.ac.jp/whois/detail/acd9a15637a319e79930f9ff86a2b16c.html