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超快单分子成像揭示了焦点粘附纳米架构和分子动力学

Takahiro K Fujiwara¹, Taka A Tsunoyama², Shinji Takeuchi³

¹Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University , Kyoto, Japan.

The Journal of cell biology

|June 6, 2023

概括

研究人员开发了一种超快的摄像头,显著加速了超分辨率显微镜 (光激活/光转换局部化显微镜和直接随机重建显微镜). 这一突破揭示了焦点粘附的动态纳米组织,揭示了一个分隔的群岛模型.

科学领域:

细胞生物学细胞生物学
显微镜的使用方法
生物物理学的生物物理.

背景情况:

像光激活/光转换局部化显微镜 (PALM) 和直接静态重建显微镜 (dSTORM) 这样的超分辨率显微镜技术对于研究细胞结构至关重要.
标准数据采集时间限制了这些方法可以实现的时空分辨率.

研究的目的:

开发和应用一个超快的摄像系统,以显著减少数据采集时间在PALM和dSTORM.
在空前的时空尺度上研究焦点粘附 (FAs) 的动态纳米组织.

主要方法:

开发一种用于增强显微镜的新型超快摄像头.
将相机应用于PALM (使用mEos3.2) 和dSTORM (使用HMSiR) 成像,实现约30倍的数据采集时间缩短.
同时的双色PALM-dSTORM和超快的 (10kHz) 单分子跟踪.

主要成果:

在更大的视野中实现了29nm (PALM) 和19nm (dSTORM) 的定位精度.
揭示了FA的动态纳米组织,从而提出了一个分隔的群岛FA模型.
在分隔的液体膜内确定了FA-蛋白岛屿 (13-100nm) (74nm区分在FA内与FA外的109nm区分).
通过蜂扩散和320nm的形成向岛屿征集综合素.

结论:

开发的超快摄像头能够在以前无法获得的时空尺度上进行高精度超分辨率成像.
分隔群岛模型为焦点粘附中的动态纳米组织和蛋白质招募机制提供了新的见解.
这一进步为研究纳米级细胞动态开辟了新的途径.

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