STED Nanoscopy Imaging of Cellular Lipid Droplets Employing a Superior Organic Fluorescent Probe

扫描电镜 荧光 共焦 荧光寿命成像显微镜 化学 受激发射 脂滴 显微镜 活体细胞成像 共焦显微镜 分子成像 光学 激光器 物理 体内 生物技术 生物 细胞 生物化学
作者
Guannan Liu,Guishan Peng,Jianan Dai,Ri Zhou,Chenguang Wang,Xu Yan,Xiaoteng Jia,Xiaomin Liu,Yuan Gao,Lijun Wang,Geyu Lu
出处
期刊:Analytical Chemistry [American Chemical Society]
卷期号:93 (44): 14784-14791 被引量:32
标识
DOI:10.1021/acs.analchem.1c03474
摘要

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.
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