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Development of an automated microscopic system for optogenetics using a mercury lamp fluorescence microscope

Research Project

Project/Area Number 24659087
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeSingle-year Grants
Research Field General anatomy (including Histology/Embryology)
Research InstitutionKagawa University

Principal Investigator

ARAKI NOBUKAZU  香川大学, 医学部, 教授 (10202748)

Co-Investigator(Kenkyū-buntansha) EGAMI Youhei  香川大学, 医学部, 助教 (80432780)
KAWAI Katsuhisa  香川大学, 医学部, 助教 (80534510)
Co-Investigator(Renkei-kenkyūsha) MIYAKE Katsuya  香川大学, 医学部, 准教授 (30219745)
Research Collaborator KATO Takuma  香川大学, 医学部, 助教 (70625673)
Project Period (FY) 2012-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2012: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywords顕微鏡技術 / 細胞・組織 / 光制御 / シグナル伝達 / 分子スイッチ / 蛍光顕微鏡 / オプトジェネティクス / 細胞 / イメージング / 光遺伝学
Research Abstract

Photomanipulation of genetically encoded photoactivatable proteins, also known as optogenetics, is one of the most innovative techniques in the fields of cell biology. Although photomanipulation is usually performed by diverting the photobleaching mode of a confocal laser microscope, photobleaching by the laser scanning unit is not always suitable for photoactivation. Therefore, we attempted to develop a simple automated wide-field fluorescence microscopy system for optogenetics. An automated fluorescence microscope can be controlled through MetaMorph imaging software, making it possible to acquire time-lapse, multiwavelength images of live cells. We wrote a macro program to change the excitation filter for photoactivation and illumination area during the intervals of image acquisition. When this program was run on the microscope, cells expressing photoactivatable Rac1 showed lamellipodial extension and cell surface ruffling in the illuminated region.

Report

(3 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • Research Products

    (19 results)

All 2014 2013 2012 Other

All Journal Article (3 results) (of which Peer Reviewed: 3 results,  Open Access: 1 results,  Acknowledgement Compliant: 1 results) Presentation (14 results) (of which Invited: 1 results) Remarks (2 results)

  • [Journal Article] Development of an automated fluorescence microscopy system for photomanipulation of genetically encoded photoactivatable proteins (optogenetics) in live cells.2014

    • Author(s)
      Araki N, Ikeda Y, Kato T, Kawai K, Egami Y, Miyake K, Tsurumaki N, Yamaguchi M.
    • Journal Title

      Microscopy (Oxf).

      Volume: 63 Issue: 3 Pages: 255-260

    • DOI

      10.1093/jmicro/dfu003

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Rac1-dependent lamellipodial motility in prostate cancer PC-3 cells revealed by optogenetic control of Rac1 activity2014

    • Author(s)
      Kato, T., Kawai, K., Egami,Y., Kahehi, Y., Araki, N.
    • Journal Title

      PLOS ONE

      Volume: (in press)

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Dissecting the roles of Rac1 activation and deactivation in macropinocytosis using microscopic photo-manipulation2013

    • Author(s)
      Fujii, M., Kawai, K., Egami, Y., Araki, N.
    • Journal Title

      Scientific reports

      Volume: 3 Issue: 1 Pages: 2385-2385

    • DOI

      10.1038/srep02385

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed
  • [Presentation] 前立腺がん細胞移動(migration)における低分子量GTPase Rac1を介した制御機構のoptogenetics解析2014

    • Author(s)
      加藤琢磨、川合克久、江上洋平、荒木伸一、筧善行
    • Organizer
      第23回泌尿器科分子・細胞研究会
    • Place of Presentation
      山形市
    • Related Report
      2013 Annual Research Report 2013 Final Research Report
  • [Presentation] Zymosan貪食過程におけるRit1 GTPaseのライブセルイメージング2014

    • Author(s)
      江上洋平、川合克久、荒木伸一
    • Organizer
      第119回日本解剖学会総会全国学術集会
    • Place of Presentation
      栃木県下野市
    • Related Report
      2013 Final Research Report
  • [Presentation] ファゴゾーム成熟過程におけるRab35の役割2014

    • Author(s)
      川合克久、江上洋平、荒木伸一
    • Organizer
      第119回日本解剖学会総会全国学術集会
    • Place of Presentation
      栃木県下野市
    • Related Report
      2013 Final Research Report
  • [Presentation] Rac1光制御により生細胞のマクロパイノサイトーシス過程を顕微鏡で操作する2013

    • Author(s)
      荒木伸一、川合克久、藤井誠、江上洋平
    • Organizer
      日本顕微鏡学会第69回学術講演会
    • Place of Presentation
      大阪府吹田市
    • Related Report
      2013 Annual Research Report 2013 Final Research Report
  • [Presentation] 水銀アーク光源蛍光顕微鏡をベースとしたphoto-manipulationシステムの構築2013

    • Author(s)
      荒木伸一、池田結香、加藤琢磨、川合克久、江上洋平、三宅克也
    • Organizer
      日本解剖学会第68回中国・四国支部学術集会
    • Place of Presentation
      鳥取県米子市
    • Related Report
      2013 Annual Research Report 2013 Final Research Report
  • [Presentation] Zymosan貪食過程におけるRit1 GTPaseの機能解析2013

    • Author(s)
      江上洋平、川合克久、荒木伸一
    • Organizer
      第54回日本組織細胞化学会総会・学術集会
    • Place of Presentation
      東京
    • Related Report
      2013 Final Research Report
  • [Presentation] 光制御Rac1を用いたマクロパイノサイトーシス過程におけるRac1活性の役割解明2012

    • Author(s)
      藤井誠, 荒木伸一
    • Organizer
      第65回薬理学会西南部会
    • Place of Presentation
      熊本
    • Year and Date
      2012-11-23
    • Related Report
      2013 Final Research Report
  • [Presentation] Photo-activatable Rac1を用いたマクロパイノサイトーシスの光制御2012

    • Author(s)
      藤井誠, 荒木伸一
    • Organizer
      第6回トランスポーター研究会九州部会
    • Place of Presentation
      福岡
    • Year and Date
      2012-09-01
    • Related Report
      2013 Final Research Report
  • [Presentation] Control of macropinocytosis by photo- manipulation of Rac12012

    • Author(s)
      Araki, N., Fujii, M., Kawai, K. and Egami, Y.
    • Organizer
      14^<th> International Congress of Histochemistry and Cytochemistry
    • Place of Presentation
      京都
    • Related Report
      2013 Final Research Report
  • [Presentation] Photo-activatable Rac1を用いてマクロパイノサイトーシスの形成・成熟過程を制御する2012

    • Author(s)
      藤井誠, 荒木伸一
    • Organizer
      第35回日本分子生物学会年会
    • Place of Presentation
      福岡
    • Related Report
      2013 Final Research Report
  • [Presentation] Control of Macropinocytosis by Photo-manipulation of Rac1 Activity.2012

    • Author(s)
      Araki, N.
    • Organizer
      14th International Congress of Histochemistry and Cytochemistry
    • Place of Presentation
      Kyoto
    • Related Report
      2012 Research-status Report
    • Invited
  • [Presentation] Photo-activatable Rac1を用いてマクロパイノサイトーシスの形成・成熟過程を制御する2012

    • Author(s)
      藤井 誠
    • Organizer
      第35回分子生物学会
    • Place of Presentation
      福岡
    • Related Report
      2012 Research-status Report
  • [Presentation] Photo-activatable Rac1を用いたマクロパイノサイトーシスの光制御

    • Author(s)
      藤井 誠
    • Organizer
      第6回 トランスポーター研究会九州部会
    • Place of Presentation
      福岡
    • Related Report
      2012 Research-status Report
  • [Presentation] 光制御Rac1を用いたマクロパイノサイトーシス過程におけるRac1活性の役割解明

    • Author(s)
      藤井 誠
    • Organizer
      第65回日本薬理学会西南部会
    • Place of Presentation
      熊本
    • Related Report
      2012 Research-status Report
  • [Remarks] マクロプログラムダウンロードサイト

    • URL

      http://jmicro.oxfordjournals.org/content/suppl/2014/01/18/dfu003.DC1/dfu003supp_data1.txt

    • Related Report
      2013 Final Research Report
  • [Remarks] マクロプログラムダウンロードサイト

    • URL

      http://jmicro.oxfordjournals.org/content/suppl/2014/01/18/dfu003.DC1/dfu003supp_data2.txt

    • Related Report
      2013 Final Research Report

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Published: 2013-05-31   Modified: 2019-07-29  

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