Development of microscope system for single fluorescent molecule imaging in vivo

• Project/Area Number: 24659092
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: General anatomy (including Histology/Embryology)
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: YASUSHI Okada 独立行政法人理化学研究所, 生命システム研究センター, チームリーダー (50272430)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 1分子イメージング / ライブイメージング / ゼブラフィッシュ / 蛍光ライブイメージング / 蛍光顕微鏡 / 共焦点顕微鏡 / in vivo イメージング

Research Abstract

Since the development of the single molecule imaging technique in Japan, it has been applied to wide areas of researches including in vitro analysis of molecular motors and single molecule measurements of signal transduction at the plasma membrane. However, this method is based on the total internal reflection microscopy, which limited its application to the region closer than 100 nm from the cover glass surface. In this project, we have tried to break this limitation and to observe single molecule deep inside the cell. By the combination of a Nipow-type confocal microscope and a high sensitivity-low noise camera, we have successfully observe the dynamics of single molecule deep inside the cell. We have further applied this technique for the super resolution imaging deep inside the cell.

Research Products

• [Journal Article] Viscosity and drag force involved in organelle transport: Investigation of the fluctuation dissipation theorem. (2013)
  • Author(s): K. Hayashi, C.G. Pack, M. K. Sato, K. Mouri, K. Kaizu, K. Takahashi and Y. Okada
  • Journal Title: The European Physical Journal E Volume: 36 Pages: 136-136
  • DOI: 10.1140/epje/i2013-13136-6
  • Peer Reviewed
• [Journal Article] Genetically encoded system to track histone modifications in vivo (2013)
  • Author(s): Yuko Sato, Masanori Mukai, Jun Ueda, Michiko Muraki, Timothy J. Stasevich, Naoki Horikoshi, Tomoya Kujirai, Hiroaki Kita, Taisuke Kimura, Seiji Hira, Yasushi Okada, Yoko Hayashi-Takanaka, Chikashi Obuse, Hitoshi Kurumizaka, Atsuo Kawahara, Kazuo Yamagata, Naohito Nozaki and Hiroshi Kimura
  • Journal Title: Scientific Rep. Volume: 3 Pages: 2436-2436
  • DOI: 10.1038/srep02436
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A spontaneously blinking fluorophore based on intramolecular spirocyclization for live-cell super-resolution imaging
  • Author(s): S. Uno, M. Kamiya, T. Yoshihara, K. Sugawara, K. Okabe, M. C. Tarham, H. Fujita, T. Funatsu, Y. Okada, S. Tobita and Y. Urano
  • Journal Title: Nat Chem Volume: (in press)
  • Peer Reviewed
• [Presentation] 高速・広視野・高分解能ライブセルイメージングのための超解像顕微鏡開発 (2014)
  • Author(s): 岡田康志
  • Organizer: 日本顕微鏡学会第70回記念学術講演会
  • Place of Presentation: 幕張
  • Year and Date: 2014-05-13
• [Presentation] 超解像顕微鏡の現状と課題～補償光学への期待 (2014)
  • Author(s): 岡田康志
  • Organizer: 若手研究者による分野間連携研究プロジェクトシンポジウムすばる望遠鏡から顕微鏡へ
  • Place of Presentation: 東京
  • Year and Date: 2014-03-24
• [Presentation] Imaging-based analyses of the intracellular navigation mechanism of molecular motor, kinesin (2013)
  • Author(s): Y. Okada
  • Organizer: 第36回日本分子生物学会シンポジウム
  • Place of Presentation: 神戸
  • Year and Date: 2013-12-03
• [Presentation] Super-resolution live imaging of intracellular transport system (2013)
  • Author(s): Y. Okada
  • Organizer: Super Imaging 2013
  • Place of Presentation: 浜松
  • Year and Date: 2013-12-02
• [Presentation] Atomic mechanism for the intracellular navigation of molecular motor, kinesin (2013)
  • Author(s): Y. Okada
  • Organizer: Joint Weizmann-MBI mechanobiology conference
  • Place of Presentation: Singapore
  • Year and Date: 2013-10-21
• [Presentation] 誘導放出制御法(STED)およびその他の手法を用いた超解像ライブイメージング (2013)
  • Author(s): 岡田康志
  • Organizer: バイオイメージング学会第22回学術集会シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2013-09-15
• [Presentation] 細胞内物質輸送の制御機構解明を目指した超解像ライブイメージング手法の開発 (2012)
  • Author(s): 岡田康志
  • Organizer: 第2回クリニカルサミット
  • Place of Presentation: 東京
  • Year and Date: 2012-12-25
• [Presentation] ライブセルイメージングのための超解像蛍光顕微鏡法 (2012)
  • Author(s): 岡田康志
  • Organizer: 日本バイオイメージング学会第21回学術集会シンポジウム
  • Place of Presentation: 京都
  • Year and Date: 2012-08-27