Isotropic super-resolution light-sheet microscopy of dynamic intracellular structures at subsecond timescales
Isotropic super-resolution light-sheet microscopy of dynamic intracellular structures at subsecond timescales
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DOI：
10.1038/s41592-022-01395-5
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发表时间：
2022-03
期刊：
影响因子：
48
通讯作者：
Yuxuan Zhao;Meng Zhang;Wenting Zhang;Yao Zhou;Longbiao Chen;Qing Liu;Peng Wang;Rong Chen;Xinxin Duan;Feifan Chen;Huan Deng;Yunfei Wei;Peng Fei;Yu-Hui Zhang
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文献类型：
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作者：
Yuxuan Zhao;Meng Zhang;Wenting Zhang;Yao Zhou;Longbiao Chen;Qing Liu;Peng Wang;Rong Chen;Xinxin Duan;Feifan Chen;Huan Deng;Yunfei Wei;Peng Fei;Yu-Hui Zhang
在整个活细胞的三维空间中，细胞内结构之间的动态相互作用的长期可视化对于更好地理解它们的功能至关重要，但由于现有的三维荧光显微镜技术的局限性，例如轴向分辨率不足，体积成像率低和光漂白，这项任务仍然具有挑战性。在这里，我们提出了一种渐进式深度学习超分辨率策略与双环调制选择性平面照明显微镜设计的结合，该设计能够以大约100 nm的各向异性空间分辨率在三维中以大约17 Hz的速度在几个小时内可视化活细胞内细胞结构的动态。利用这种方法，我们揭示了活细胞中内质网（ER）和线粒体之间复杂的空间关系和相互作用，为内质网介导的线粒体分裂提供了新的见解。我们还研究了参与线粒体裂变的Drp1寡聚物的运动，并在三维空间上揭示了Drp1与线粒体之间的动态相互作用。
Long-term visualization of the dynamic interactions between intracellular structures throughout the three-dimensional space of whole live cells is essential to better understand their functions, but this task remains challenging due to the limitations of existing three-dimensional fluorescence microscopy techniques, such as an insufficient axial resolution, low volumetric imaging rate and photobleaching. Here, we present the combination of a progressive deep-learning super-resolution strategy with a double-ring-modulated selective plane illumination microscopy design capable of visualizing the dynamics of intracellular structures in live cells for hours at an isotropic spatial resolution of roughly 100 nm in three dimensions at speeds up to roughly 17 Hz. Using this approach, we reveal the complex spatial relationships and interactions between endoplasmic reticulum (ER) and mitochondria throughout live cells, providing new insights into ER-mediated mitochondrial division. We also examined the motion of Drp1 oligomers involved in mitochondrial fission and revealed the dynamic interactions between Drp1 and mitochondria in three dimensions.