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Establishing Systems to Obtain Uniform and Stable Cell Population Based on Massively Parallel Manipulation of Single Cells

Research Project

Project/Area Number 16H06074
Research Category

Grant-in-Aid for Young Scientists (A)

Allocation TypeSingle-year Grants
Research Field Intelligent mechanics/Mechanical systems
Research InstitutionToyohashi University of Technology

Principal Investigator

Nagai Moeto  豊橋技術科学大学, 工学(系)研究科(研究院), 講師 (00580557)

Research Collaborator Santra Tuhin Subhra  
Chiou Pei-Yu (Eric)  
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥25,220,000 (Direct Cost: ¥19,400,000、Indirect Cost: ¥5,820,000)
Fiscal Year 2018: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2017: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
Fiscal Year 2016: ¥13,390,000 (Direct Cost: ¥10,300,000、Indirect Cost: ¥3,090,000)
Keywords超並列単一細胞操作 / ナノ秒レーザ / 誘電泳動プローブアレイ / 誘電泳動力 / レーザ誘起衝撃波 / 光照射 / DMD / 光硬化性ゲル / マイクロコンタクトプリンティング / 単一細胞操作 / 細胞加工 / 超並列 / 核内デリバリー / 多点光照射 / レーザーアブレーション / 衝撃波 / 細胞内デリバリ / マイクロシステム / 単一細胞 / 細胞操作 / パルスレーザ / 細胞内デリバリー / 誘電泳動 / 3次元電極 / マイクロ・ナノデバイス / 知能機械 / バイオ関連機器 / ナノバイオ / 生物・生体工学
Outline of Final Research Achievements

We have advanced the development of the massively parallel intranuclear delivery technology , and we established an optical system necessary for cell poration. Cells were cultured on a Ti substrate and irradiated with nanosecond pulsed laser to generate shock waves by photo-ablation. With the developed optical system, we carried out a cell viability assay and time lapse observation of irradiated cells.
The research team developed dielectrophoretic probe array as a cell pickup tool successfully. We have developed an electrode wiring process to control individual probes individually and a fabrication process of a three-dimensional probe array for cell manipulation into microwells. Parallel cell manipulation was demonstrated by generating dielectrophoretic forces.

Academic Significance and Societal Importance of the Research Achievements

開発した光照射光学系と光穿孔法は,低侵襲かつ高い効率でのトランスフェクションにつながる.さらに誘電泳動プローブアレイは,狙った細胞の選別につながる.これらの開発した技術を統合し,時系列で発現が安定な細胞を構築する技術が創出される.微細加工技術を利用した構造作製ならびにマイクロ・ナノメカトロニクス技術を元にした操作・観察を複合活用して,複数の単一細胞を処理して医学・生物学に貢献する新しい学術基盤が構築できる.
本研究の技術・システムは,基礎的な細胞構築方法を提供し,遺伝子組換え細胞を利用した広い科学技術と産業を活性化する.バイオ医薬品,iPS細胞を用いた再生医療・細胞医療の社会での普及に寄与する.

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Annual Research Report
  • 2016 Annual Research Report
  • Research Products

    (30 results)

All 2019 2018 2017 2016 Other

All Int'l Joint Research (2 results) Journal Article (8 results) (of which Open Access: 4 results,  Peer Reviewed: 2 results,  Acknowledgement Compliant: 1 results) Presentation (14 results) (of which Int'l Joint Research: 4 results,  Invited: 2 results) Book (1 results) Remarks (5 results)

  • [Int'l Joint Research] インド工科大学マドラス校(インド)

    • Related Report
      2018 Annual Research Report
  • [Int'l Joint Research] インド工科大学マドラス校(インド)

    • Related Report
      2017 Annual Research Report
  • [Journal Article] Scalable hollow nanoneedle array using stepper lithography for parallel intracellular delivery2019

    • Author(s)
      Nagai Moeto、Miyamoto Tokuma、Hizawa Takeshi、Shibata Takayuki
    • Journal Title

      Precision Engineering

      Volume: 55 Pages: 439-446

    • DOI

      10.1016/j.precisioneng.2018.10.014

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Phototactic Algae-Driven Unidirectional Transport of Submillimeter-Sized Cargo in a Microchannel2019

    • Author(s)
      Nagai Moeto、Hirano Takahiro、Shibata Takayuki
    • Journal Title

      Micromachines

      Volume: 10 Issue: 2 Pages: 130-130

    • DOI

      10.3390/mi10020130

    • Related Report
      2018 Annual Research Report
    • Open Access
  • [Journal Article] Batch Assembly of SU-8 Movable Components in Channel Under Mild Conditions for Dynamic Microsystems:Application to Biohybrid Systems2019

    • Author(s)
      Nagai Moeto、Tanizaki Kohei、Shibata Takayuki
    • Journal Title

      Journal of Microelectromechanical Systems

      Volume: In press Issue: 3 Pages: 1-10

    • DOI

      10.1109/jmems.2019.2907285

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Current Trends of Microfluidic Single-Cell Technologies2018

    • Author(s)
      Shinde Pallavi、Mohan Loganathan、Kumar Amogh、Dey Koyel、Maddi Anjali、Patananan Alexander、Tseng Fan-Gang、Chang Hwan-You、Nagai Moeto、Santra Tuhin
    • Journal Title

      International Journal of Molecular Sciences

      Volume: 19 Issue: 10 Pages: 3143-3143

    • DOI

      10.3390/ijms19103143

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Rapid prototyping of PDMS microchannels for animal and plant cells using cutting plotter and double casting2018

    • Author(s)
      NAGAI Moeto、TADA Kyohei、SHIBATA Takayuki
    • Journal Title

      Mechanical Engineering Letters

      Volume: 4 Issue: 0 Pages: 18-00377-18-00377

    • DOI

      10.1299/mel.18-00377

    • NAID

      130007537457

    • ISSN
      2189-5236
    • Related Report
      2018 Annual Research Report
    • Open Access
  • [Journal Article] Single-cell electroporation: current trends, applications and future prospects2018

    • Author(s)
      Kar Srabani、Loganathan Mohan、Dey Koyel、Shinde Pallavi、Chang Hwan-You、Nagai Moeto、Santra Tuhin Subhra
    • Journal Title

      Journal of Micromechanics and Microengineering

      Volume: 28 Issue: 12 Pages: 123002-123002

    • DOI

      10.1088/1361-6439/aae5ae

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Pick-and-Place Operation of Single Cell Using Optical and Electrical Measurements for Robust Manipulation2017

    • Author(s)
      Moeto Nagai, Keita Kato, Kiyotaka Oohara, Takayuki Shibata
    • Journal Title

      Micromachines

      Volume: 8 Issue: 12 Pages: 350-350

    • DOI

      10.3390/mi8120350

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Minimally Invasive Intracellular Delivery Based on Electrokinetic Forces Combined with Vibration-Assisted Cell Membrane Perforation2017

    • Author(s)
      T. Shibata, T. Ozawa, Y. Ito, K. Yamamoto, M. Nagai
    • Journal Title

      Japanese Journal of Applied Physics

      Volume: 56 Issue: 1 Pages: 017001-017001

    • DOI

      10.7567/jjap.56.017001

    • NAID

      210000147356

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] Parallelized Single-cell Optoporation Using Nanosecond Laser for Homogeneous Transfection2019

    • Author(s)
      Harsh Gupta, Naoki Iwasaki, Shin Sawai, Pallavi Shinde, Tuhin Subhra Santra, Takayuki Shibata, Moeto Nagai
    • Organizer
      2019年度精密工学会春季大会学術講演会,pp. 331-332,東京電機大学 北千住キャンパス
    • Related Report
      2018 Annual Research Report
  • [Presentation] マイクロ・ナノ医療機器の創出 各要素技術の整理と実用化の可能性検討2019

    • Author(s)
      永井萌土
    • Organizer
      株式会社情報機構セミナー[東京・大井町] きゅりあん
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 超並列細胞内デリバリーシステム開発に向けた照射光学系の評価及びデバイス作製2019

    • Author(s)
      澤井 慎, Gupta Harsh, Santra Tuhin, 鹿毛あずさ,柴田隆行,永井萌土
    • Organizer
      令和元年度電気学会E部門総合研究会
    • Related Report
      2018 Annual Research Report
  • [Presentation] ナノ秒パルスレーザ用いた超並列核内デリバリ技術の開発―光照射による細胞膜穿孔の実証―2018

    • Author(s)
      舟橋圭佑,澤井慎,Gupta Harsh, Shinde Pallavi, Santra Tuhin, 柴田 隆行, 永井 萌土
    • Organizer
      精密工学会2018年度春季大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] パルス光照射で生成するマイクロバブルを用いた位置選択的な細胞膜穿孔技術の開発2017

    • Author(s)
      舟橋 圭佑,Tuhin Santra,永井 萌土,柴田 隆行
    • Organizer
      ロボティクス・メカトロニクス講演会2017
    • Place of Presentation
      ビッグパレットふくしま,福島県郡山市
    • Year and Date
      2017-05-10
    • Related Report
      2016 Annual Research Report
  • [Presentation] Single cell processing microsystems for high throughput function analysis of cells2017

    • Author(s)
      Moeto NAGAI
    • Organizer
      IEEE NEMS 2017
    • Place of Presentation
      University of California, Los Angeles
    • Year and Date
      2017-04-09
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] ナノ秒パルス光を用いた超並列核内デリバリー技術の開発  ―流路開口部の作製技術と光学系の構築―2017

    • Author(s)
      舟橋圭佑,サントラツーヒン,永井萌土,柴田隆行
    • Organizer
      精密工学会 2017年度春季大会
    • Place of Presentation
      慶應義塾大学矢上キャンパス
    • Year and Date
      2017-03-13
    • Related Report
      2016 Annual Research Report
  • [Presentation] 超並列単一細胞操作のための誘電泳動プローブアレイの開発2017

    • Author(s)
      坂本良作,佐伯拓朗,永井萌土,柴田隆行
    • Organizer
      精密工学会 2017年度春季大会
    • Place of Presentation
      慶應義塾大学矢上キャンパス
    • Year and Date
      2017-03-13
    • Related Report
      2016 Annual Research Report
  • [Presentation] 超並列単一細胞加工:基盤技術と集積システムの開発2017

    • Author(s)
      永井萌土
    • Organizer
      定量生物学の会 第八回年会
    • Place of Presentation
      自然科学研究機構 岡崎カンファレンスセンター (OCC),愛知県岡崎市
    • Year and Date
      2017-01-08
    • Related Report
      2016 Annual Research Report
  • [Presentation] Optical Transfection System using Pulse Laser for Massively Parallel Localized Intracellular Delivery2017

    • Author(s)
      Keisuke Funahashi, Shin Sawai, Tuhin Subhra Santra, Takayuki Shibata, and Moeto Nagai
    • Organizer
      IEEE MHS 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 超並列の細胞核内デリバリのためのレーザ誘起プラズマ発生デバイスの開発2017

    • Author(s)
      舟橋圭佑,澤井 慎,Tuhin Santra,永井萌土,柴田隆行
    • Organizer
      第8回マイクロ・ナノ工学シンポジウム
    • Related Report
      2017 Annual Research Report
  • [Presentation] 大面積細胞パターニングのための空圧マイクロコンタクトプリント技術の開発2017

    • Author(s)
      岩崎 真己, 峰村 俊輝, 永井 萌土, 柴田 隆行
    • Organizer
      第8回マイクロ・ナノ工学シンポジウム
    • Related Report
      2017 Annual Research Report
  • [Presentation] 3D Manipulation of Single Cells Using Dielectrophoretic Probe Array2016

    • Author(s)
      Ryosaku Sakamoto, Takuro Saeki, Moeto Nagai, and Takayuki Shibata
    • Organizer
      IEEE MHS 2016
    • Place of Presentation
      Nagoya University, Noyori Confernece Hall
    • Year and Date
      2016-11-28
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Massively Parallel Localized Intracellular Delivery for Uniform Transfection Simple Fabrication Process of Multilayer Microdevice2016

    • Author(s)
      Keisuke Funahashi, Moeto Nagai, Tuhin S. Santra, and Takayuki Shibata
    • Organizer
      The Irago Conference 2016
    • Place of Presentation
      Auditorium, University of Electro-Communications, Chofu
    • Year and Date
      2016-11-01
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Book] Handbook of Single Cell Technologies, Mechanoporation: Toward Single Cell Approaches2018

    • Author(s)
      Amogh KumarL. MohanPallavi ShindeHwan-You ChangMoeto NagaiTuhin Subhra Santra
    • Total Pages
      29
    • Publisher
      Springer Nature
    • Related Report
      2018 Annual Research Report
  • [Remarks] 豊橋技術科学大学 機械工学系 マイクロ・ナノ機械システム研究室

    • URL

      http://mems.me.tut.ac.jp/

    • Related Report
      2018 Annual Research Report
  • [Remarks] バイオマイクロシステムの研究 永井萌土

    • URL

      https://sites.google.com/site/moeton/

    • Related Report
      2018 Annual Research Report
  • [Remarks] バイオマイクロシステムの研究

    • URL

      https://sites.google.com/site/moeton/home

    • Related Report
      2017 Annual Research Report
  • [Remarks] 研究者「永井萌土」に関するウェブページ

    • URL

      https://sites.google.com/site/moeton/

    • Related Report
      2016 Annual Research Report
  • [Remarks] 大学での研究に関するウェブページ

    • URL

      https://www.tut.ac.jp/university/faculty/me/667.html

    • Related Report
      2016 Annual Research Report

URL: 

Published: 2016-04-21   Modified: 2020-03-30  

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