Chemical Sciences Division, Saha Institute of Nuclear Physics , 1/AF Bidhannagar, Kolkata 700064, India.
Anal Chem. 2014 Mar 4;86(5):2740-6. doi: 10.1021/ac404160v. Epub 2014 Feb 21.
We introduce a new rhodamine-rhodanine-based "turn-on" fluorescent sensor (RR1) and describe its application for detection of mercury, including in solution, in live cells, and in a living vertebrate organism. The sensor RR1, which is a one-pot synthesis from rhodamine B, undergoes a rapid and irreversible 1:1 stoichiometric reaction with Hg(2+) in aqueous medium. Using fluorescence correlation spectroscopy (FCS), RR1 was shown to detect the presence of as low as a 0.5 pM concentration of Hg(2+). It may also lend itself to tagging with biomolecules and nanoparticles, leading to the possibility of organelle-specific Hg detection. Results of experiments with mammalian cells and zebrafish show that RR1 is cell and organism permeable and that it responds selectively to mercury ions over other metal ions. In addition, real-time monitoring of inorganic mercury ion uptake by cells and live zebrafish using this chemosensor shows that saturation of mercury ion uptake occurs within 20-30 min in cells and organisms. We also demonstrate the acquisition of high-resolution real-time distribution maps of inorganic mercury (Hg(2+)) in the zebrafish brain by using a simple fluorescence confocal imaging technique.
我们介绍了一种新型的基于罗丹明-硫代罗丹明的“开启”荧光传感器(RR1),并描述了其在汞检测方面的应用,包括在溶液中、活细胞中和活体脊椎动物中。RR1 传感器可由罗丹明 B 一锅合成,在水相介质中与 Hg(2+)发生快速且不可逆的 1:1 计量比反应。荧光相关光谱(FCS)表明,RR1 可检测低至 0.5 pM 浓度的 Hg(2+)。RR1 还可能与生物分子和纳米粒子标记相结合,从而实现细胞器特异性汞检测。与哺乳动物细胞和斑马鱼的实验结果表明,RR1 可穿透细胞和生物体,并且对汞离子具有选择性响应,而对其他金属离子没有响应。此外,使用该化学传感器实时监测细胞和活体斑马鱼对无机汞离子的摄取表明,细胞和生物体中汞离子摄取的饱和在 20-30 分钟内发生。我们还通过使用简单的荧光共焦成像技术,证明了在斑马鱼大脑中获取无机汞(Hg(2+))的高分辨率实时分布图谱的能力。