College of Materials Science & Engineering, South China University of Technology, Guangzhou 510640, China.
Anal Chim Acta. 2012 Jul 13;734:69-78. doi: 10.1016/j.aca.2012.05.020. Epub 2012 May 23.
This article describes the design and preparation of a novel fluorescence resonance energy transfer (FRET)-based ratiometric sensor with the polymer nanoparticle as scaffold for detecting Hg(2+) in aqueous media. In this study, a fluorescent dye fluorescein isothiocyanate (FITC, served as the donor) and a spirolactam rhodamine derivative (SRHB, served as mercury ion probe) were covalently attached onto polyethylenimine (PEI) and polyacrylic acid (PAA) respectively; and a ratiometric sensing system was then formed through the deposition of the donor- and probe-containing polyelectrolytes onto the negatively charged polymer particles via the layer-by-layer approach. The ratiometric fluorescent signal change of the system is based on the intra-particle fluorescence resonance energy transfer (FRET) process modulated by mercury ions. Under optimized structural and experimental conditions, the particle-based detection system exhibits stable response for Hg(2+) in aqueous media. More importantly, in this newly developed particle-based detection system formed by LBL approach, varied numbers of the PAA/PEI layers which served as the spacer could be placed between the donor-containing layer and the probe-containing layer, hence the donor-acceptor distance and energy transfer efficiency could be effectively tuned (from ca. 25% to 76%), this approach has well solved the problem for many particle-based FRET systems that the donor-acceptor distance cannot be precisely controlled. Also, it is found that the ratiometric sensor is applicable in a pH range of 4.6-7.3 in water with the detection limit of 200 nM. This approach may provide a new strategy for ratiometric detection of analytes in some environmental and biological applications.
本文描述了一种新型荧光共振能量转移(FRET)比率传感器的设计和制备,该传感器以聚合物纳米颗粒为支架,用于在水介质中检测 Hg(2+)。在本研究中,将荧光染料异硫氰酸荧光素(FITC,作为供体)和螺环缩醛罗丹明衍生物(SRHB,作为汞离子探针)分别共价连接到聚乙烯亚胺(PEI)和聚丙烯酸(PAA)上;然后,通过层层自组装的方法,将含有供体和探针的聚电解质沉积到带负电荷的聚合物颗粒上,形成比率传感体系。该体系的比率荧光信号变化基于汞离子调制的颗粒内荧光共振能量转移(FRET)过程。在优化的结构和实验条件下,该基于颗粒的检测体系在水介质中对 Hg(2+)表现出稳定的响应。更重要的是,在这种通过 LBL 方法形成的新开发的基于颗粒的检测系统中,可以在含有供体的层和含有探针的层之间放置不同数量的 PAA/PEI 层作为间隔物,从而有效调节供体-受体距离和能量转移效率(从约 25%到 76%),这种方法很好地解决了许多基于颗粒的 FRET 系统中供体-受体距离无法精确控制的问题。此外,还发现该比率传感器适用于 pH 值为 4.6-7.3 的水中,检测限为 200 nM。该方法可能为在某些环境和生物应用中分析物的比率检测提供一种新策略。