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2020 年度 実施状況報告書

Mechanical stability of microtubules investigated under unidirectional strain using combined fluorescence microscopy and high-speed atomic force microscopy

研究課題

研究課題/領域番号 20K03889
研究機関大学共同利用機関法人自然科学研究機構(新分野創成センター、アストロバイオロジーセンター、生命創成探究

研究代表者

Ganser Christian  大学共同利用機関法人自然科学研究機構(新分野創成センター、アストロバイオロジーセンター、生命創成探究, 生命創成探究センター, 特任助教 (50846095)

研究期間 (年度) 2020-04-01 – 2023-03-31
キーワードhigh-speed AFM / microtubules / mechanical properties / correlated imaging
研究実績の概要

A basic mechanism that allows stretching of the substrate was successfully tested and it was confirmed that nanometer scale effects could be observed. Furthermore, the sensitivity of the atomic force microscope was improved significantly to allow for undisturbed long-time observation of fragile samples such as microtubules. It was successfully demonstrated that the tip-scan high-speed atomic force microscope can take images simultaneously with the fluorescence microscope and that both images are spatially and temporally correlated.
The force mapping of the high-speed atomic force microscope was improved so that the invasiveness was minimized further to allow undisturbed mechanical mapping of microtubules under strain.

現在までの達成度 (区分)
現在までの達成度 (区分)

2: おおむね順調に進展している

理由

The initial state of the tip-scan high-speed atomic force microscope system allowed for imaging microtubules, however, the invasiveness was considerable and caused microtubules to depolymerize during force mapping as well as during imaging. To overcome this problem, the system has been improved by optimizing the alignment of the read-out laser, including rigorous blocking of reflected light with an optical isolator. The scanner has been balanced to remove unwanted resonance frequencies in order to scan faster and more stable. In addition the optical fluorescence microscope has been fully synchronized with the atomic force microscope which finally allowed correlated observations.
All these improvements lead to a solid basis for continued research on strained microtubules.

今後の研究の推進方策

In the next steps, the mechanical properties of microtubules under different conditions (strain, nucleotide state, stabilizing agents) will be investigated using the improved combined atomic force/fluorescence microscope.
Further along the timeline, the mechanism to stretch the substrate will be improved to eventually allow in-situ straining while observing with the high-speed atomic force microscope. This will require a complex tracking algorithm to consistently follow the region of interest during strain application.

  • 研究成果

    (3件)

すべて 2020

すべて 学会発表 (3件) (うち国際学会 1件、 招待講演 1件)

  • [学会発表] High-speed atomic force microscopy based force mapping and multiscale observation in combination with fluorescence microscopy2020

    • 著者名/発表者名
      Christian Ganser
    • 学会等名
      3rd ExCELLS retreat for young researchers
  • [学会発表] High-speed atomic force microscopy as a versatile tool to study dynamical and mechanical properties of proteins2020

    • 著者名/発表者名
      Christian Ganser, Kimitoshi Takeda, Ryota Iino, Koichi Kato, Takayuki Uchihashi
    • 学会等名
      The 58th Annual Meeting of the Biophysical Society of Japan
    • 国際学会 / 招待講演
  • [学会発表] Multimodal high-speed atomic force microscopy to investigate nano-scalemechanical properties and dynamics on the protein and cell level2020

    • 著者名/発表者名
      Shigetaka Nishiguchi, Christian Ganser
    • 学会等名
      3rd ExCELLS Symposium

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公開日: 2021-12-27  

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