线粒体是细胞中最重要的细胞器之一，在信号传递和生死存亡中发挥关键作用，因此线粒体质量控制对细胞稳态维持至关重要。线粒体衍生小泡（MDVs）是近年来新发现的应对线粒体内活性氧过多引起损伤的新机制，而MDVs是如何产生、成熟、以及靶向下游细胞器等过程仍存在许多争议和疑问，其中的重要原因是MDVs的尺寸为70~150 纳米，传统显微成像难以准确连续追踪其动态过程。因此申请人将采用自主搭建的掠入射结构光超分辨显微镜(GI-SIM)在活细胞状态下对MDVs动态过程连续观测，明确其产生、成熟和靶向其它细胞器等过程。目前，申请人已经发现了MDVs从线粒体上“拉丝-断裂”的产生方式，并将继续通过结合分离纯化、质谱、高通量筛选等方法进一步研究MDVs产生的分子机制、阐明MDVs靶向选择性、寻找MDVs与其他细胞器的互作新现象等。最后，明确MDVs通路缺陷所引起的表型也为后续探索MDVs的生理功能奠定基础。

Mitochondrion is one of the most important intracellular organelles that plays critical role in signal transmission, cell survival and death. Therefore, mitochondrial quality control is of critical importance to homeostasis of the cell. Mitochondrial-derived vesicles (MDVs) is a novel mechanism that deals with the damages caused by excessive reactive oxygen species in the mitochondria. However, it still remains unknown how MDVs are generated, matured, and targeted to the downstream organelles. An obvious reason is that its size range from 70-150 nm, which is hardly to be precisely resolved and tracked by conventional fluorescence microscopy. Here, we employed the home-built grazing incidence structured illumination microscopy (GI-SIM) to clearly resolve and continually track the dynamics of MDVs in live cells, so as to gain insights into its generation, maturation, and destination. So far, we have captured the generation dynamics of MDVs in a pull-and-break manner. In addition to super-resolution imaging, our following investigations will integrate biochemical purification, mass spectrometry, and high-throughput screening to explore MDVs related molecular mechanism, targeting selectivity, and possible interactions with other organelles. Finally, to clear the phenotype caused by loss of function of MDVs-related genes, which may lay the foundation for study of physiologic function.