National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
Cell. 2018 Nov 15;175(5):1430-1442.e17. doi: 10.1016/j.cell.2018.09.057. Epub 2018 Oct 25.
In eukaryotic cells, organelles and the cytoskeleton undergo highly dynamic yet organized interactions capable of orchestrating complex cellular functions. Visualizing these interactions requires noninvasive, long-duration imaging of the intracellular environment at high spatiotemporal resolution and low background. To achieve these normally opposing goals, we developed grazing incidence structured illumination microscopy (GI-SIM) that is capable of imaging dynamic events near the basal cell cortex at 97-nm resolution and 266 frames/s over thousands of time points. We employed multi-color GI-SIM to characterize the fast dynamic interactions of diverse organelles and the cytoskeleton, shedding new light on the complex behaviors of these structures. Precise measurements of microtubule growth or shrinkage events helped distinguish among models of microtubule dynamic instability. Analysis of endoplasmic reticulum (ER) interactions with other organelles or microtubules uncovered new ER remodeling mechanisms, such as hitchhiking of the ER on motile organelles. Finally, ER-mitochondria contact sites were found to promote both mitochondrial fission and fusion.
在真核细胞中，细胞器和细胞骨架经历高度动态但组织有序的相互作用，能够协调复杂的细胞功能。可视化这些相互作用需要在高时空分辨率和低背景下对细胞内环境进行非侵入性、长时间的成像。为了实现这些通常相互矛盾的目标，我们开发了掠入射结构光照明显微镜（GI-SIM），能够以 97nm 的分辨率和 266 帧/秒的速度在数千个时间点上对基底细胞皮质附近的动态事件进行成像。我们采用多色 GI-SIM 来描述不同细胞器和细胞骨架的快速动态相互作用，为这些结构的复杂行为提供了新的认识。对微管生长或收缩事件的精确测量有助于区分微管动态不稳定性的模型。内质网（ER）与其他细胞器或微管相互作用的分析揭示了新的 ER 重塑机制，例如 ER 在运动细胞器上的搭便车。最后，发现 ER-线粒体接触位点促进线粒体裂变和融合。
