Imaging myosin-driven stress fiber contraction with molecular resolution by high-speed AFM

通过高速 AFM 以分子分辨率对肌球蛋白驱动的应力纤维收缩进行成像

• 批准号: 20H03218
• 负责人: CLEMENS・MARTIN FRANZ
• 金额: $ 11.56万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2020
• 资助国家: 日本
• 起止时间: 2020-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-20H03218/
• 关键词: HS-AFM; myosin; actin; AFM; Myosin; Actin; High-speed AFM; Bioimaging; SPM; cytoskeleton; Stress Fibers

The overall aim of the project is to image large intracellular protein assemblies, such as actin-myosin fibers, with molecular resolution under physiological conditions by high-speed atomic force microscopy (HS-AFM). However, previous HS-AFM scanners ususally have only small scan sizes. In this project phase we developed an ultrawide HS-AFM sample-scanner system able to record large topographic images (40 x 40 micrometer) containing up to 16 megapixels, providing molecular resolution throughout the image frame. With this unique scanner design we are now able for the first time to image even large intracellular structures and entire organelles in a single AFM scan with near molecular resolution aiding the quantitative analysis of such structurally heterogenous samples.

该项目的总体目标是通过高速原子力显微镜（HS-AFM）在生理条件下对大的细胞内蛋白质组装体（如肌动蛋白-肌球蛋白纤维）进行分子分辨率成像。然而，以前的HS-AFM扫描仪通常只有很小的扫描尺寸。在这个项目阶段，我们开发了一个超宽HS-AFM样品扫描仪系统，能够记录包含高达1600万像素的大型地形图像（40 × 40微米），在整个图像帧中提供分子分辨率。有了这种独特的扫描仪设计，我们现在能够第一次成像，甚至大的细胞内结构和整个细胞器在一个单一的原子力显微镜扫描与近分子分辨率，帮助定量分析这种结构异质性样品。

项目成果

Applications for HS-AFM in Cell Biology

HS-AFM 在细胞生物学中的应用

• 发表时间: 2021
• 作者: C. Furian;N. Chongdar;P. Gupta;W. Lubitz;H. Ogata;J. N. Blaza;J. A. Birrell;原幸大;Eiichiro Mori.;Clemens M. Franz
• 通讯作者: Clemens M. Franz

Atomic reconstruction of biomolecular structures from AFM images and quantitative validation of experimental data using simulated AFM scanning

利用 AFM 图像对生物分子结构进行原子重建，并使用模拟 AFM 扫描对实验数据进行定量验证

• DOI: 10.1101/2021.06.27.450070
• 发表时间: 2021
• 期刊: bioRxiv
• 作者: Amyot Romain;Marchesi Arin;Franz Clemens M;Casuso Ignacio;Flechsig Holger
• 通讯作者: Flechsig Holger

Imaging Myosin II-driven Stress Fiber Contraction in De-roofed Cells by High-Speed AFM

通过高速 AFM 对去顶细胞中肌球蛋白 II 驱动的应力纤维收缩进行成像

• 发表时间: 2020
• 作者: Nakanishi Hanayo;Hayashida Kenichi;Nishizawa Tomohiro;Oshima Atsunori;Abe Kazuhiro;Clemens M. Franz
• 通讯作者: Clemens M. Franz