Extended-resolution structured illumination imaging of endocytic and cytoskeletal dynamics

分类： 生物学 >> 生物物理学 提交时间： 2016-05-12

Li, Dong Shao, Lin Chen, Bi-Chang Betzig, Eric Zhang, Xi Zhang, Mingshu Xu, Pingyong Zhang, Xi Zhang, Mingshu Xu, Pingyong Zhang, Xi Moses, Brian Milkie, Daniel E. Beach, Jordan R. Hammer, John A., III Baird, Michelle A. Pasham, Mithun Kirchhausen, Tomas Pasham, Mithun Kirchhausen, Tomas

摘要： Super-resolution fluorescence microscopy is distinct among nanoscale imaging tools in its ability to image protein dynamics in living cells. Structured illumination microscopy (SIM) stands out in this regard because of its high speed and low illumination intensities, but typically offers only a twofold resolution gain. We extended the resolution of live-cell SIM through two approaches: ultrahigh numerical aperture SIM at 84-nanometer lateral resolution for more than 100 multicolor frames, and nonlinear SIM with patterned activation at 45- to 62-nanometer resolution for approximately 20 to 40 frames. We applied these approaches to image dynamics near the plasma membrane of spatially resolved assemblies of clathrin and caveolin, Rab5a in early endosomes, and a-actinin, often in relationship to cortical actin. In addition, we examined mitochondria, actin, and the Golgi apparatus dynamics in three dimensions.

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