State Key Laboratory of Integrated Optoelectronics, Key Laboratory of Advanced Gas Sensors of Jilin Province, College of Electronic Science & Engineering, Jilin University, Changchun 130012, People's Republic of China.
State Key Laboratory of Luminescence and Applications, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun 130033, People's Republic of China.
Anal Chem. 2021 Nov 9;93(44):14784-14791. doi: 10.1021/acs.analchem.1c03474. Epub 2021 Oct 27.
Lipid droplets (LDs) are spherical organelles that participate in numerous biological processes. In order to visualize LDs on the nanoscale, nanoscopy fluorescence imaging is considered as the most attractive technique but is substantially limited by the characteristics of fluorescent probes. Thus, the development of a superior fluorescent probe that is capable of nanoscopy fluorescence imaging has attracted enormous attention. Herein, a benzodithiophene-tetraoxide-based molecule Lipi-BDTO has been developed that can easily undergo the stimulated emission depletion (STED) process and displays high photostability. These two characteristics of fluorescent probes finely satisfy the requirements of STED nanoscopy imaging. Indeed, applying the probe for STED imaging achieves a high resolution of 65 nm, belonging to one of the leading results of LDs fluorescence imaging. Furthermore, the high photostability of this fluorescent probe enables it to monitor the dynamics of LDs by time-lapse STED imaging as well as to visualize the three-dimensional (3D) spatial distribution of LDs by 3D STED imaging. Notably, the resolution of the 3D STED image represents one of the best LDs fluorescence imaging results so far. Besides STED nanoscopy imaging, the superior utility of this fluorescent probe has been also demonstrated in two-color 3D confocal imaging and four-color confocal imaging.
脂滴（LDs）是参与众多生物过程的球形细胞器。为了在纳米尺度上可视化 LD，纳米荧光显微镜成像被认为是最有吸引力的技术，但受到荧光探针特性的极大限制。因此，开发一种能够进行纳米荧光显微镜成像的优越荧光探针引起了极大关注。在此，开发了一种基于苯并二噻吩四氧化物的分子 Lipi-BDTO，它可以很容易地经历受激发射耗竭（STED）过程，并显示出高的光稳定性。荧光探针的这两个特性很好地满足了 STED 纳米显微镜成像的要求。实际上，应用该探针进行 STED 成像可实现 65nm 的高分辨率，属于 LD 荧光成像的领先结果之一。此外，该荧光探针的高光稳定性使其能够通过时间分辨 STED 成像来监测 LD 的动力学，以及通过 3D STED 成像来可视化 LD 的三维（3D）空间分布。值得注意的是，3D STED 图像的分辨率代表了迄今为止 LD 荧光成像的最佳结果之一。除了 STED 纳米显微镜成像外，该荧光探针的优越性能还在双色 3D 共聚焦成像和四色共聚焦成像中得到了证明。
