Development of real-time secretion analysis of immune single cells

• Project/Area Number: 23770189
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biophysics
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: SHIRASAKI Yoshitaka 独立行政法人理化学研究所, 免疫ゲノミクス研究グループ, 研究員 (70469948)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2011: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
• Keywords: 単一細胞解析 / イムノアッセイ / 分泌測定 / マイクロウェル / CYTOP / 単一細胞測定 / 免疫システム

Research Abstract

We developed a platform for real-time secretion analysis at the single cell resolution. The platform enabled to monitor secretion dynamics of thousand of single cells with 30-min time resolution for over 8 hours. We also demonstrated multi-parametric measurement of secretion dynamics and cellular states with 1-min time resolution. We applied our platform to inflammatory responses of immune cells and observed intercellular and intracellular heterogeneity of cytokine secretion.

Research Products

• [Journal Article] Toward an understanding of immune cell sociology: real-time monitoring of cytokine secretion at the single-cell level (2012)
  • Author(s): Shirasaki Y, Yamagishi M, Shimura N, Hijikata A,Ohara O.
  • Journal Title: IUBMB Life Volume: 65 Issue: 5 Pages: 28-34
  • DOI: 10.1002/iub.1175
  • Peer Reviewed
• [Journal Article] Fabrication of high-aspect-ratio amorphous perfluorinated polymer structure for total in ternal reflection fluorescence microscopy (2011)
  • Author(s): A.Nakahara, Y.Shirasaki, K.Kawai, O.Ohara, J.Mizuno
  • Journal Title: Microelectronic Engineering Volume: 88 Issue: 8 Pages: 1817-1820
  • DOI: 10.1016/j.mee.2011.02.117
  • Peer Reviewed
• [Journal Article] Microfluidic active sorting of DNA molecules labeled with single quantum dots using flow switching by a hydrogel sol-gel transition (2011)
  • Author(s): Mai Haneoka
  • Journal Title: Sensors and Actuators B : Chemical Volume: 159 Issue: 1 Pages: 314-320
  • DOI: 10.1016/j.snb.2011.06.043
  • Peer Reviewed
• [Journal Article] Single cell real time secretion assay using amorphous fluoropolymer microwell array (2011)
  • Author(s): Shirasaki Y., Nakahara A., Shimura N., Yamagishi M., Mizuno J., Ohara O. and Shoji S.
  • Journal Title: Solid-State Sensors, Actuators and Microsystems Conference (TRANSDUCERS), 2011 16th International technical digest Pages: 755-758
  • DOI: 10.1109/transducers.2011.5969330
  • Peer Reviewed
• [Journal Article] 微細凹版法を用いたタンパク質高分泌株の高効率単一細胞スクリーニング (2011)
  • Author(s): 白崎 善隆
  • Journal Title: ファルマシア Volume: 47 Pages: 1163-1163
  • NAID: 110009884886
• [Journal Article] Single cell analysis of the proinflammatory responses of mast cell by a real time secretion assay (2011)
  • Author(s): Shirasaki Y., Nakahara A., Shimura N., Suzuki N., Yamagishi M., Mizuno J., Shoji S. and Ohara O.
  • Journal Title: microTAS 2011 technical program Volume: - Pages: 843-845
  • Peer Reviewed
• [Journal Article] Microfluidic active sorting of DNA molecules labeled with single quantum dots using flow switching by a hydrogel sol-gel transition (2011)
  • Author(s): Haneoka M., Shirasaki Y., Sugino H., Sekiguchi T., Yoon D. H., Iizuka R., Shoji S. and Funatsu T.
  • Journal Title: microTAS 2011 technical program Volume: - Pages: 1773-1775
  • Peer Reviewed
• [Journal Article] 微細凹版法を用いたタンパク質高分泌株の高効率単一細胞スクリーニング (2011)
  • Author(s): 白崎善隆
  • Journal Title: ファルマシア Volume: 47 Pages: 1163-1163
  • NAID: 110009884886
• [Presentation] 単一細胞イムノアッセイによるNLRP3 体細胞モザイクの機能的解析の試み (2013)
  • Author(s): 白崎 善隆
  • Organizer: 第6回日本免疫不全症研究会
  • Place of Presentation: 品川インターシティホール, 東京都
  • Year and Date: 2013-01-26
• [Presentation] Measurement of inframmatory cytokine secretion from human monocytes after inframasome activation (2012)
  • Author(s): Y.Shirasaki
  • Organizer: The 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences
  • Place of Presentation: OKINAWA, JAPAN
• [Presentation] マクロファージ細胞株におけるLPS 活性化時の遺伝子発現の細胞間ゆらぎに細胞間相互作用が与える影響 (2012)
  • Author(s): 山岸 舞
  • Organizer: 第50回生物物理学会年会
  • Place of Presentation: 名古屋大学,愛知県
• [Presentation] 炎症時におけるヒト単球サイトカイン分泌のライブセルイメージング (2012)
  • Author(s): 白崎 善隆
  • Organizer: 第50回生物物理学会年会
  • Place of Presentation: 名古屋大学,愛知県
• [Presentation] マクロファージ細胞株における LPS 活性化時の遺伝子発現の細胞間ゆらぎに細胞間相互作用が与える影響 (2012)
  • Author(s): M. Yamagishi, Y. Shirasaki, N. Shimura, N. Suzuki, O. Ohara
  • Organizer: 第50回生物物理学会年会
  • Place of Presentation: 名古屋大学,愛知県
• [Presentation] Microfluidic active sorting of DNA molecules labeled with single quantum dots using flow switching by a hydrogel sol-gel transition (2011)
  • Author(s): Haneoka M.
  • Organizer: The 15th international conference on miniaturized systems for chemistry and life sciences
  • Place of Presentation: Seattle (U.S.A.)
  • Year and Date: 2011-10-05
• [Presentation] Single cell analysis of the proinflammatory responses of mast cell by a real time secretion assay (2011)
  • Author(s): Shirasaki Y.
  • Organizer: The 15th international conference on miniaturized systems for chemistry and life sciences
  • Place of Presentation: Seattle (U.S.A.)
  • Year and Date: 2011-10-04
• [Presentation] Single cell real time secretion assay using amorphous fluoropolymer microwell array (2011)
  • Author(s): Nakahara A.
  • Organizer: The 16th international conference on Solid-State Sensors, Actuators and Microsystems (TRANSDUCERS'11)
  • Place of Presentation: Beijing, China
  • Year and Date: 2011-06-06
• [Presentation] Single cell real time secretion assay using amorphous fluoropolymer microwell array (2011)
  • Author(s): Shirasaki Y., Nakahara A., Shimura N., Yamagishi M., Mizuno J., Ohara O. and Shoji S.
  • Organizer: The 16th international conference on Solid-State Sensors, Actuators and Microsystems (TRANSDUCERS’11)
  • Place of Presentation: Beijing (China)