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Development of near-field optical microscopy for hydrogen ion concentration and membrane potential using environmental sensitivity dye

Research Project

Project/Area Number 22310078
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Nanomaterials/Nanobioscience
Research InstitutionTokyo University of Agriculture and Technology

Principal Investigator

UMEDA Norihiro  東京農工大学, 大学院・工学研究院, 教授 (60111803)

Co-Investigator(Kenkyū-buntansha) IWAMI Kentarou  東京農工大学, 大学院・工学研究院, 准教授 (80514710)
OHTA Yoshihiro  東京農工大学, 大学院・工学研究院, 准教授 (10223843)
Project Period (FY) 2010 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥19,760,000 (Direct Cost: ¥15,200,000、Indirect Cost: ¥4,560,000)
Fiscal Year 2012: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2011: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2010: ¥13,650,000 (Direct Cost: ¥10,500,000、Indirect Cost: ¥3,150,000)
Keywords近接場光学顕微鏡 / 蛍光感受性色素 / ミトコンドリア / pH / ネクローシス / 多光子吸収 / プロトンポンプ / 蛍光色素 / pH / フェムト秒レーザー
Research Abstract

In this study, a dual-wavelength pH-sensitive dye was excited by two-photon absorption initiated using afemtosecond pulse laser. In addition, fluorescence from the dye was directly collected from the fluorescent point using the collection-mode probe of a scanning near-field optical microscope. By thisproposed method, a pH calibration curve was obtained from the fluorescent intensity ratio of the dyesolution, and temporal pH variations with 0.1 s time resolution following the addition of acid wereobserved. Moreover, mitochondrial activity on the basis of the pH changes was successfully observed in three different mitochondrial densities.

Report

(4 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • Research Products

    (34 results)

All 2013 2012 2011 2010 Other

All Journal Article (6 results) (of which Peer Reviewed: 4 results) Presentation (24 results) Remarks (4 results)

  • [Journal Article] pH measurement using dual wavelength fluorescent ratio by two-photon excitation for mitochondrial activity2012

    • Author(s)
      Y. Kanazashi, Y. Li, T. Onojima, K. Iwami, Y. Ohta, N. Umeda
    • Journal Title

      Jpn. J. Appl. Phys

      Volume: 51 Issue: 11R Pages: 117011-117015

    • DOI

      10.1143/jjap.51.117001

    • NAID

      40019496267

    • Related Report
      2012 Annual Research Report 2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] ミトコンドリアの内部密度の評価法の開発とその応用2012

    • Author(s)
      長谷田圭亮、兼松啓太、太田善浩
    • Journal Title

      バイオイメージング

      Volume: 21 Pages: 15-21

    • NAID

      10031161857

    • URL

      http://www.tuat.ac.jp/~ohta

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Observation of Mitochondrial Activity Based on Temporal and Spatial pH Variation Measured by Near-Field Fluorescent Ratiometry2011

    • Author(s)
      Yongbo Li, Ryosuke Shinohara, Kentaro Iwami, Yoshihiro Ohta a n d Norihiro Umeda
    • Journal Title

      Science China Physics, Mechanics and Astronomy

      Volume: 54 Issue: 12 Pages: 2225-2229

    • DOI

      10.1007/s11433-011-4533-4

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Two-photon fluorescence near-field pH measurement for mitochondria activity2011

    • Author(s)
      Y. Kanazashi, Y. Li, T. Onojima, K. Iwami, Y. Ohta, N. Umeda
    • Journal Title

      SPIE Digital Library NanoScience + Engineering: Instrumentation, Metrology, and Standards for Nanomanufacturing, Optics, and Semiconductors V

      Volume: 81050E

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Two-photon fluorescence near-field pH measurement for mitochondria activity2011

    • Author(s)
      Y.Kanazashi, Y.Li, T.Onojima, K.Iwami, Y.Ohta, N.Umeda
    • Journal Title

      SPIE Digital Library NanoScience + Engineering

    • Related Report
      2011 Annual Research Report
  • [Journal Article] Dual Wavelength Fluorescent Ratiometric pH Measurement by Scanning Near-Field Optical Microscopy2010

    • Author(s)
      Y.Li, R.Shinohara, K.Iwami, Y.Ohta, N.Umeda
    • Journal Title

      Proceedings of SPIE

      Volume: 7544

    • Related Report
      2010 Annual Research Report
  • [Presentation] Evaluation of Mitochondrial Activity by Membrane Potential Observation and pH measurement2013

    • Author(s)
      Y. Kanazashi, Y. Nomura, D. Yoshimatsu, K. Iwami, Y. Ohta, N. Umeda
    • Organizer
      APNFO2013
    • Place of Presentation
      Singapore
    • Year and Date
      2013-07-05
    • Related Report
      2012 Final Research Report
  • [Presentation] ミトコンドリア活性評価のための膜電位観測と2光子吸収pH測定2013

    • Author(s)
      金指康明,野村雄一,李永波,吉松大輝,岩見健太郎,太田善浩,梅田倫弘
    • Organizer
      2013年春季第60回応用物理学関係連合講演
    • Place of Presentation
      厚木、神奈川工科大学
    • Year and Date
      2013-03-28
    • Related Report
      2012 Final Research Report
  • [Presentation] ミトコンドリア活性評価のための膜電位観測と2 光子吸収pH 測定2013

    • Author(s)
      金指 康明, 野村 雄一, 李 永波, 吉松 大輝, 岩見 健太郎, 太田 善浩, 梅田 倫弘
    • Organizer
      2013 年春季第60 回応用物理学関係連合講演会
    • Place of Presentation
      厚木・神奈川工科大学
    • Related Report
      2012 Annual Research Report
  • [Presentation] 膜電位観測と2光子吸収励起pH測定によるミトコンドリア活性評価2012

    • Author(s)
      野村雄一,金指康明,吉松大輝,岩見健太郎,太田善浩,梅田倫弘
    • Organizer
      日本光学会年次学術講演会
    • Place of Presentation
      東京・タワーホール船越
    • Year and Date
      2012-10-25
    • Related Report
      2012 Final Research Report
  • [Presentation] pH measurement for mitochondrial activity evaluation using two-photon excitation2012

    • Author(s)
      Y. Kanazashi, Y. Li, T. Onojima, K. Iwami, Y. Ohta, N. Umeda
    • Organizer
      NFO12
    • Place of Presentation
      San Sebastian, Spain
    • Year and Date
      2012-09-06
    • Related Report
      2012 Final Research Report
  • [Presentation] pH measurement for mitochondrial activity evaluation using fluorescent ratiometry by two-photon absorption2012

    • Author(s)
      Y. Kanazashi, Y. Li, T. Onojima, K. Iwami, Y. Ohta, N. Umeda
    • Organizer
      ICHC2012
    • Place of Presentation
      京都・京都国際会館
    • Year and Date
      2012-08-28
    • Related Report
      2012 Annual Research Report 2012 Final Research Report
  • [Presentation] ミトコンドリア活性評価のための2光子吸収励起による近接場pH測定2012

    • Author(s)
      金指康明,李永波,小野島匠,岩見健太郎,太田善浩,梅田倫弘
    • Organizer
      日本光学会・ナノオプティクス研究グループ研究討論会第20回記念シンポジウム
    • Place of Presentation
      横浜、慶応大学
    • Year and Date
      2012-05-23
    • Related Report
      2012 Final Research Report
  • [Presentation] 2光子吸収励起によるpH値測定を用いたミトコンドリア活性評価2012

    • Author(s)
      金指康明,李永波,小野島匠,岩見健太郎,太田善浩,梅田倫弘
    • Organizer
      2012年春季第59回応用物理学関係連合講演
    • Place of Presentation
      東京、早稲田大学
    • Year and Date
      2012-03-17
    • Related Report
      2012 Final Research Report
  • [Presentation] 2光子吸収励起によるpH値測定を用いたミトコンドリア活性評価2012

    • Author(s)
      金指康明, 李永波, 小野島匠, 岩見健太郎, 太田善浩, 梅田倫弘
    • Organizer
      第59回応用物理学関係連合講演
    • Place of Presentation
      早稲田大学
    • Year and Date
      2012-03-17
    • Related Report
      2011 Annual Research Report
  • [Presentation] ミトコンドリア活性評価のための2 光子吸収励起による近接場pH 測定2012

    • Author(s)
      金指 康明, 李 永波, 小野島 匠, 岩見 健太郎, 太田 善浩, 梅田 倫弘
    • Organizer
      日本光学会・ナノオプティクス研究グループ研究討論会第20回記念シンポジウム
    • Place of Presentation
      横浜・慶応大学
    • Related Report
      2012 Annual Research Report
  • [Presentation] 膜電位観測と2光子吸収励起pH測定によるミトコンドリア活性評価2012

    • Author(s)
      野村 雄一, 金指 康明, 吉松 大輝, 岩見 健太郎, 太田 善浩, 梅田 倫弘
    • Organizer
      日本光学会年次学術講演会OPJ2012
    • Place of Presentation
      東京・タワーホール船越
    • Related Report
      2012 Annual Research Report
  • [Presentation] pH measurement for mitochondrial activity evaluation using two-photon excitation2012

    • Author(s)
      Y. Kanazashi, Y. Li, T. Onojima, K. Iwami, Y. Ohta, N. Umeda
    • Organizer
      NFO12
    • Place of Presentation
      San Sebastian/Spain
    • Related Report
      2012 Annual Research Report
  • [Presentation] Two-photon fluorescence near-field pH measurement for mitochondria activity2011

    • Author(s)
      Y. Kanazashi, Y. Li, T. Onojima, K. Iwami, Y. Ohta, N. Umeda
    • Organizer
      SPIE Optics + Photonics
    • Place of Presentation
      San Diego, USA
    • Year and Date
      2011-08-25
    • Related Report
      2012 Final Research Report
  • [Presentation] 2光子励起による2波長蛍光レシオを用いた微小領域pH測定2011

    • Author(s)
      金指康明,李永波,小野島匠,岩見健太郎,太田善浩,梅田倫弘
    • Organizer
      第37回レーザ顕微鏡研究会
    • Place of Presentation
      和光、理化学研究所
    • Year and Date
      2011-07-06
    • Related Report
      2012 Final Research Report
  • [Presentation] 2光子励起による2波長蛍光レシオを用いた微小領域pH測定2011

    • Author(s)
      金指康明, 李永波, 小野島匠, 岩見健太郎, 太田善浩, 梅田倫弘
    • Organizer
      第37回レーザ顕微鏡研究会
    • Place of Presentation
      理化学研究所(和光)
    • Year and Date
      2011-07-06
    • Related Report
      2011 Annual Research Report
  • [Presentation] Two-photon fluorescence near-field pH measurement for mitochondria activity2011

    • Author(s)
      Y.Kanazashi, Y.Li, T.Onojima, K.Iwami, Y.Ohta, N.Umeda
    • Organizer
      SPIE Optics + Photonics
    • Place of Presentation
      San Diego (USA)
    • Year and Date
      2011-05-25
    • Related Report
      2011 Annual Research Report
  • [Presentation] 2光子蛍光励起による近接場pH値測定2011

    • Author(s)
      金指康明,李永波,小野島匠,岩見健太郎,太田善浩,梅田倫弘
    • Organizer
      2011年春季第58回応用物理学関係連合講演
    • Place of Presentation
      厚木、神奈川工科大学
    • Year and Date
      2011-03-27
    • Related Report
      2012 Final Research Report
  • [Presentation] Temporal and Spatial pH Variation Measurement by Near-Field Fluorescent Ratiometry for Observation of Mitochondrial Activity2010

    • Author(s)
      Y. Li, Y. Kanazashi, K. Iwami, Y. Ohta, N. Umeda
    • Organizer
      東京農工大学・電気通信大学第7回合同シンポジウム"ナノ未来材料とコヒーレント光科学"
    • Place of Presentation
      調布、電気通信大学
    • Year and Date
      2010-12-11
    • Related Report
      2012 Final Research Report
  • [Presentation] 蛍光レシオ近接場顕微鏡法によるミトコンドリア活性の観測2010

    • Author(s)
      李永波,金指康明,岩見健太郎,太田善浩,梅田倫弘
    • Organizer
      日本光学会年次学術講演会
    • Place of Presentation
      東京、中央大学
    • Year and Date
      2010-11-10
    • Related Report
      2012 Final Research Report
  • [Presentation] 蛍光レシオ近接場顕微鏡法によるミトコンドリア活性の観測2010

    • Author(s)
      李永波、金指康明、岩見健太郎、太田善浩、梅田倫弘
    • Organizer
      日本光学会年次学術講演会Optics, Photonics Japan 2010
    • Place of Presentation
      中央大学駿河台記念館、東京都
    • Year and Date
      2010-11-10
    • Related Report
      2010 Annual Research Report
  • [Presentation] 近接場光学プローブを備えた蛍光顕微鏡によるミトコンドリア周囲のpHの計測の試み2010

    • Author(s)
      長谷田圭亮,李永波,篠原亮輔,金指康明,小野島匠,岩見健太郎,梅田倫弘,太田善浩
    • Organizer
      第48回日本生物物理学会年会
    • Place of Presentation
      仙台、東北大学
    • Year and Date
      2010-09-20
    • Related Report
      2012 Final Research Report
  • [Presentation] Scanning Near-field Optical Microscope for pH measurement using dual wavelength fluorescent spectroscopy2010

    • Author(s)
      Y.Li, R.Shinohara, K.Iwami, Y.Ohta, N.Umeda
    • Organizer
      11th International Conference on Near-field Optics, Nanophotonics, Related Techniques (NFO-11)
    • Place of Presentation
      Peking University, Beijing, China
    • Year and Date
      2010-08-31
    • Related Report
      2010 Annual Research Report
  • [Presentation] Dual Wavelength Fluorescent Ratiometric pH Measurement by Scanning Near-Field Optical Microscopy2010

    • Author(s)
      Y.Li, R.Shinohara, K.Iwami, Y.Ohta, N.Umeda
    • Organizer
      6^<th> International Symposium on Precision Engineering Measurements and Instrumentation (ISPEMI 2010)
    • Place of Presentation
      Hangzhou, China
    • Year and Date
      2010-08-10
    • Related Report
      2010 Annual Research Report
  • [Presentation] 蛍光レシオ近接場顕微鏡法によるミトコンドリア活性の観測2010

    • Author(s)
      李永波,金指康明,岩見健太郎,太田善浩,梅田倫弘
    • Organizer
      第36回レーザ顕微鏡研究会
    • Place of Presentation
      和光、理化学研究所
    • Year and Date
      2010-07-06
    • Related Report
      2012 Final Research Report
  • [Remarks]

    • URL

      http://www.tuat.ac.jp/~umedalab

    • Related Report
      2012 Final Research Report
  • [Remarks] 東京農工大学工学府梅田研究室

    • URL

      http://www.tuat.ac.jp/~umedalab

    • Related Report
      2012 Annual Research Report
  • [Remarks]

    • URL

      http://www.tuat.ac.jp/~umedalab

    • Related Report
      2011 Annual Research Report
  • [Remarks]

    • URL

      http://www.tuat.ac.jp/~umedalab/

    • Related Report
      2010 Annual Research Report

URL: 

Published: 2010-08-23   Modified: 2019-07-29  

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