A Phosphole Oxide Based Fluorescent Dye with Exceptional Resistance to Photobleaching: A Practical Tool for Continuous Imaging in STED Microscopy
作者:Chenguang Wang、Aiko Fukazawa、Masayasu Taki、Yoshikatsu Sato、Tetsuya Higashiyama、Shigehiro Yamaguchi
DOI:10.1002/anie.201507939
日期:2015.12.7
depletion (STED) microscopy represented a major breakthrough in cellular and molecular biology. However, the intense laser beams required for both excitation and STED usually provoke rapid photobleaching of fluorescent molecular probes, which significantly limits the performance and practical utility of STED microscopy. We herein developed a photoresistant fluorescent dye C‐Naphox as a practical tool for
受激发射损耗(STED)显微镜的发展代表了细胞和分子生物学的重大突破。然而,激发和STED所需的强激光束通常会引起荧光分子探针的快速光致漂白,这大大限制了STED显微镜的性能和实用性。我们在此开发了一种耐光荧光染料C-Naphox作为STED成像的实用工具。与使用一个激发λ = 405或488nm的在质子溶剂激光,C-Naphox表现出强烈的红色/橙色荧光(量子产率Φ ˚F > 0.7)具有大的斯托克斯位移(大约5900厘米-1)。即使使用Xe灯(300W,在照射后λ EX= 460 nm,半峰全宽(FWHM)= 11 nm)持续12小时,C-Naphox保持99.5%完好无损。C-Naphox的高光阻允许对HeLa细胞进行重复STED成像。即使记录了50张STED图像,初始荧光强度的83%仍然存在。