对细胞器互作进行三维（3D）超分辨水平动态检测将突破现有绝大多数研究在二维检测上的局限，对进一步理解和揭示细胞器的生理功能具有重要意义。晶格光片超分辨显微成像技术为3D超分辨水平活细胞动态检测提供了一种有力工具，并已成为当前研究热点之一。但是要利用其实现3D超分辨水平细胞器互作动态检测，现有细胞器荧光探针在荧光亮度和抗光漂白性能方面还远不能满足其需求并已成为了关键瓶颈之一。本申请项目拟基于独有新型活细胞超分辨有机荧光探针构建技术，通过进一步改造有机荧光探针结构及相关活细胞探针投递技术，得到一系列能在活细胞内特异性标记各种细胞器并具有高荧光亮度、强抗光漂白能力，能进行3D超分辨水平细胞器互作动态检测的有机荧光探针。这些荧光探针将为3D超分辨水平细胞器互作动态研究提供强有力的工具。在此基础上，将进行3D超分辨水平细胞器互作及互作网络的动态研究，从而在3D水平发现新的细胞器互作方式及互作网络。

More and more studies have indicated that intracellular organelles are integrated into dynamic networks and collaborate on various cellular tasks rather than acting as isolated entities. There are various dynamic interactions between organelles, which occur not only in small sites, but also in three-dimensional (3D) space. However, most of the existing studies are restricted to two-dimensional level, which greatly limits the understanding of organelle interactions and their dynamic networks. .Recently developed lattice light-sheet super-resolution microscopy (LSM) not only can achieve a spatial resolution beyond the diffraction limit but also can perform 3D live-cell imaging with fast imaging speed and low photo-toxicity. Therefore, LSM provides a powerful approach for visualizing dynamic processes in live cells at a 3D super-resolution level. However, the applications of LSM for characterizing the dynamic interactions and the dynamic networks among organelles remain challenging due to limitations of the existing fluorescent probes, such as low fluorescence intensity and poor photo-stability..In this project, we will develop a series of novel cell-permeable organic fluorescent probes for organelles with high fluorescence intensity and excellent photo-stability by optimizing the structures of organic probes and the sequences of cytosolic delivery peptide vehicles. Using our probes and LSM, we will record the dynamic interactions and the dynamic networks between organelles at a 3D super-resolution level, which will reveal new organelle interaction modes. Our fluorescent probes herein also will provide a new avenue for understanding the functions and the dynamic interplay between organelles in live cells at 3D level.