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Development of high-throughput Real-time single-cell secretion assay systems

Research Project

Project/Area Number 25440074
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Biophysics
Research InstitutionThe University of Tokyo (2014-2015)
Institute of Physical and Chemical Research (2013)

Principal Investigator

Shirasaki Yoshitaka  東京大学, 理学(系)研究科(研究院), 助教 (70469948)

Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywords1細胞生物学 / サイトカイン分泌 / 全反射顕微鏡 / 導波路照明 / 1細胞計測 / 細胞分泌 / 全反射照明 / 単一細胞解析 / 分泌活性測定 / 全反射蛍光顕微鏡 / サイトカイン / イムノアッセイ
Outline of Final Research Achievements

This project focused on developing the techniques for the single-cell secretion dynamics measurement, which will contribute to understanding of the detailed mechanism of Immune systems reguration at single-cell resolution. During this project, I succeeded in developing a novel micro fabricated waveguide chip for wide field total internal reflection fluorescence microscopy (TIRFM).
This chip has the optical refraction index matching layer on both side of quartz light wave guide slide, and nano-litter well array made by PDMS on top side.Using this chip with the wave-guide illuminator, we measured single-cell cytokine secretion dynamics from hundreds of single-cells every minute for over 2hours.
This optical wave-guide based TIRFM system has great potential for revealing single-cell secretion dynamics and will contribute to develop the measurement technique for analyzing cell-cell interaction via humoral factors.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • 2013 Research-status Report
  • Research Products

    (11 results)

All 2015 2014 2013

All Journal Article (3 results) (of which Peer Reviewed: 2 results,  Open Access: 2 results,  Acknowledgement Compliant: 1 results) Presentation (8 results)

  • [Journal Article] 1細胞分泌実時間イメージング法2015

    • Author(s)
      白崎善隆、山岸舞、小原收、上村想太郎
    • Journal Title

      細胞工学

      Volume: 34(3) Pages: 304-309

    • NAID

      40020377011

    • Related Report
      2014 Research-status Report
  • [Journal Article] Single-Cell Imaging of Caspase-1 Dynamics Reveals an All-or-None Inflammasome Signaling Response.2014

    • Author(s)
      Liu, T., Yamaguchi, Y., Shirasaki, Y., Shikada, K., Yamagishi, M., Hoshino, K., Kaisho, T., Takemoto, K., Suzuki, T., Kuranaga, E., Ohara, O., Miura, M.
    • Journal Title

      Cell Reports

      Volume: 8 Issue: 4 Pages: 974-982

    • DOI

      10.1016/j.celrep.2014.07.012

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Real-time single-cell imaging of protein secretion2014

    • Author(s)
      Yamagishi, M. Suzuki, N. Izawa, K. Nakahara, A. Mizuno, J. Shoji, S. Heike, T. Harada, Y. Nishikomori, R. Ohara, O.
    • Journal Title

      Sci Rep

      Volume: 4 Issue: 1 Pages: 4736-4736

    • DOI

      10.1038/srep04736

    • Related Report
      2014 Research-status Report 2013 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Real-time single-cell secretion imaging revealed heterogeneity of the cell secretion2015

    • Author(s)
      Yoshitaka Shirasaki, Ting Liu, Yoshifumi Yamaguchi, Mai Yamagishi, Nobutake Suzuki, Masayuki Miura, Osamu Ohara, Sotaro Uemura
    • Organizer
      The 53rd Annual Meeting of the Biophysical Society of Japan
    • Place of Presentation
      金沢大学 角間キャンパス (石川県金沢市)
    • Year and Date
      2015-09-13
    • Related Report
      2015 Annual Research Report
  • [Presentation] HIGH-THROUGHPUT SINGLE-CELL SECRETION MESUREMENT ON AN OPTICAL WAVEGUDE CHIP2014

    • Author(s)
      Yoshitaka Shirasaki, Nobutake Suzuki, Mai Yamagishi, Asahi Nakahara, Shuichi Shoji and Osamu Ohara
    • Organizer
      The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences
    • Place of Presentation
      テキサス、サンアントニオ
    • Year and Date
      2014-10-26 – 2014-10-30
    • Related Report
      2014 Research-status Report
  • [Presentation] 1細胞分泌実時間測定によるIL-1β非古典的分泌機序の解明2014

    • Author(s)
      白崎 善隆、劉 霆、山口 良文、山岸 舞、鈴木 信勇、井澤 和司、水野 潤、庄子 習一、原田 慶恵、西小森 隆太、平家 俊男、三浦 正幸、小原 收
    • Organizer
      第52回日本生物物理学会年会
    • Place of Presentation
      北海道、札幌コンベンションセンター
    • Year and Date
      2014-09-25 – 2014-09-27
    • Related Report
      2014 Research-status Report
  • [Presentation] Time-resolved live-cell FluoroSpot assay on Total Internal Reflection Fluorescence Microscopy2014

    • Author(s)
      Yoshitaka Shirasaki, Nobutake Suzuki, Mai Yamagishi, Osamu Ohara
    • Organizer
      EMBL Conference Series Microfluidics 2014
    • Place of Presentation
      ドイツ、ハイデルベルグ
    • Year and Date
      2014-07-23 – 2014-07-25
    • Related Report
      2014 Research-status Report
  • [Presentation] 炎症性細胞死に伴うIL-1βサイトカイン分泌の1細胞イメージング2014

    • Author(s)
      白崎 善隆、劉 霆、山口 良文、山岸 舞、鈴木 信勇、三浦 正幸、小原 收
    • Organizer
      第23回日本Cell Death学会学術集会
    • Place of Presentation
      東京都、東京医科歯科大学
    • Year and Date
      2014-07-18 – 2014-07-19
    • Related Report
      2014 Research-status Report
  • [Presentation] リアルタイム分泌イメージング法を用いたインフラマソーム活性化に伴うIL-1β分泌機序の解析2014

    • Author(s)
      白崎 善隆, 山岸 舞, 鈴木 信勇, 劉 霆, 山口 良文, 改正 恒康, 星野 克明, 三浦 正幸, 小原 收
    • Organizer
      第66回日本細胞生物学会大会
    • Place of Presentation
      奈良県、新公会堂
    • Year and Date
      2014-06-11 – 2014-06-13
    • Related Report
      2014 Research-status Report
  • [Presentation] Real-time single-cell imaging of IL-1β secretion by Inflammasomes2014

    • Author(s)
      Yoshitaka Shirasaki, Mai Yamagishi, Kazushi Izawa, Hirotsugu Oda, Toshio Heike, Ryuta Nishikomori, Osamu Ohara
    • Organizer
      CSH 2014 meeting on Gene expression & signaling in the immune system
    • Place of Presentation
      ニューヨーク、コールドスプリングハーバーラボラトリー
    • Year and Date
      2014-04-22 – 2014-04-26
    • Related Report
      2014 Research-status Report
  • [Presentation] Real-time secretion analysis 64 revealed correlation of IL-1beta release and loss of cell membrane integrity2013

    • Author(s)
      Shirasaki Y, Yamagishi M, Izawa K, Nakagawa K, Nakahara A, Suzuki N, Mizuno J, Sekiguchi T, Heike T, Nishikomori R, Shoji S, Ohara O
    • Organizer
      MicroTAS2013
    • Place of Presentation
      Freiburg, Germany
    • Related Report
      2013 Research-status Report

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Published: 2014-07-25   Modified: 2019-07-29  

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