超高分辨成像技术使我们可以在纳米尺度观察目标蛋白在细胞中的精确定位，是蛋白质研究和荧光成像领域国际前沿的研究热点和发展趋势。然而，受限于复杂的成像设备、荧光探针以及难以同时具有高的时间和空间分辨率，活细胞超高分辨成像技术仍面临诸多挑战。本课题将基于前期自主研发的基于单分子定位的贝叶斯成像方法SIMBA，发展多种新型活细胞超高分辨成像技术及其对应的荧光蛋白探针，并拓展它们在细胞内膜动态结构观察等生物学中的应用。具体包括发展更高时空分辨率成像技术qSIMBA，双色活细胞成像技术dSIMBA，超高时空分辨率蛋白质-蛋白质相关作用成像技术PPI-SIMBA，以及单分子跟踪超高分辨成像技术smtSIMBA。本项目的实施将极大地促进当前活细胞超高分辨成像技术的发展及其在蛋白质定位和转运研究、蛋白质转录和翻译等诸多领域中的广泛应用。

Super-resolution (SR) imaging technologies enable us to define the accurate localization of labeled protein in cells and have been hot issues in recent years in popular imaging field. However, because the current SR methods either require sophisticated optical setups, specific fluorescent probes and deep experts or have difficulties to achieve high spatial and temporal resolutions simultaneously, the applications of live-cell SR microscopy have remained technically challenging. Based on our recently developed live-cell SIMBA microscopy, we will develop several novel SR techniques including: quick-SIMBA with higher temporal-spatial resolution, dual color SIMBA technique, PPI-SIMBA for detecting protein-protein interaction and the single molecular tracking-SIMBA technique. The novel SR techniques in this project will greatly increase the development of live-cell SR microscopies and their applications in the process of protein trafficking, transcription, and translation.