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Development of liquid chamber for pulsed electron microscopy

Research Project

Project/Area Number 19K22384
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
Research InstitutionNagoya University

Principal Investigator

Narita Akihiro  名古屋大学, 理学研究科, 准教授 (30360613)

Project Period (FY) 2019-06-28 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2020: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2019: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords電子顕微鏡 / 蛋白質動態 / パルス電子顕微鏡 / 溶液構造 / タンパク質動態 / 液中観察 / 蛋白質 / ダイナミクス
Outline of Research at the Start

開発中のパルス電子顕微鏡を用いると1ミリ秒以下の露光時間で一枚の電子顕微鏡写真が撮れる。パルス間隔を開けることでダメージを抑えながら長時間動態観察することもできる。本研究では、このパルス電子顕微鏡によって液中の蛋白質動態を直接観察するための液体試料観察法の開発を行う。より具体的には、1: 電子顕微鏡観察に向いた溶液条件の探索、2:従来よりコントラストの高い溶液チャンバー作成法の開発を行う。

Outline of Final Research Achievements

In this study, we developed a liquid chamber for observing protein dynamics with a pulsed electron microscope. We made a chip with a 40 nm SiN (silicon nitride) film stuck on silicon with holes, and made a liquid holder by sticking two of these chips together. As a result of trial and error, it was finally possible to obtain a contrast in which the movement of colloidal gold with a diameter of 20 nm in a solution could be constantly observed with a normal electron microscope (not a pulsed electron microscope). From the comparison with the commercially available solution chamber, the solution thickness seems to be less than 100 nm, and the success rate of solution observation exceeds 50%, almost reaching the practical level.

Academic Significance and Societal Importance of the Research Achievements

パルス電子顕微鏡による溶液状態の観察が可能になれば、蛋白質動態だけでなく、リポソームやミセル、エマルジョンのような溶液構造についての微細動態観察も可能になり、生物学だけでなく、医薬、食品、化粧品、塗料など応用分野は広い。本研究はまだ完成にまでは至っていないが、必要条件を満たす溶液チャンバーの作成にほぼ成功しており、重要な一歩を刻むことができたと考えている。

Report

(3 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • Research Products

    (5 results)

All 2020 2019

All Presentation (5 results) (of which Int'l Joint Research: 2 results,  Invited: 5 results)

  • [Presentation] アクチン線維構造と運動マシナリー2020

    • Author(s)
      成田哲博
    • Organizer
      第43回分子生物学会年会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] 細胞運動のしくみ解明に挑戦する構造生物学 ~アクチン線維の精密解析~2020

    • Author(s)
      成田哲博
    • Organizer
      未来に挑戦する構造生物科学
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] フォトカソード電子顕微鏡に生物分野から期待できること2020

    • Author(s)
      成田哲博
    • Organizer
      第238回有機エレクトロニクス研究会
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] High resolution structural analysis of the actin filaments and actin related proteins2019

    • Author(s)
      Akihiro Narita
    • Organizer
      Frontiers in Cellular, Viral and Molecular Microscopy with Cryo-specimen Preparation Techniques, 英国ブリストル大学
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Cofilin functions and regulation from a structural view point,2019

    • Author(s)
      Akihiro Narita
    • Organizer
      The 11th Toyota Riken International Workshop “Actin Filament: beyond the atomic resolution structures”
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research / Invited

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Published: 2019-07-04   Modified: 2022-01-27  

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