Unité de Pharmacologie Chimique et Génétique et Imagerie, CNRS 8151, École Nationale Supérieure de Chimie de Paris, Chimie ParisTech, Université Paris Descartes, Paris, France.
Lab Chip. 2011 Apr 7;11(7):1342-50. doi: 10.1039/c0lc00585a. Epub 2011 Feb 14.
In this work we report on the design, microfabrication and analytical performances of a new electrochemical sensor array (ESA) which allows for the first time the simultaneous amperometric detection of nitric oxide (NO) and peroxynitrite (ONOO(-)), two biologically relevant molecules. The on-chip device includes individually addressable sets of gold ultramicroelectrodes (UMEs) of 50 µm diameter, Ag/AgCl reference electrode and gold counter electrode. The electrodes are separated into two groups; each has one reference electrode, one counter electrode and 110 UMEs specifically tailored to detect a specific analyte. The ESA is incorporated on a custom interface with a cell culture well and spring contact pins that can be easily interconnected to an external multichannel potentiostat. Each UME of the network dedicated to the detection of NO is electrochemically modified by electrodepositing thin layers of poly(eugenol) and poly(phenol). The detection of NO is performed amperometrically at 0.8 V vs. Ag/AgCl in phosphate buffer solution (PBS, pH = 7.4) and other buffers adapted to biological cell culture, using a NO-donor. The network of UMEs dedicated to the detection of ONOO(-) is used without further chemical modification of the surface and the uncoated gold electrodes operate at -0.1 V vs. Ag/AgCl to detect the reduction of ONOOH in PBS. The selectivity issue of both sensors against major biologically relevant interfering analytes is examined. Simultaneous detection of NO and ONOO(-) in PBS is also achieved.
在这项工作中，我们报告了一种新的电化学传感器阵列（ESA）的设计、微制造和分析性能，该阵列首次允许同时安培检测两种具有生物学相关性的分子——一氧化氮（NO）和过氧亚硝酸盐（ONOO(-)）。该芯片装置包括可单独寻址的 50 µm 直径的金超微电极（UME）、Ag/AgCl 参比电极和金对电极的集合。这些电极被分为两组；每组都有一个参比电极、一个对电极和 110 个 UME，专门用于检测特定的分析物。ESA 被集成到一个带有细胞培养池和弹簧接触销的定制接口上，可以很容易地与外部多通道电位计进行互连。专门用于检测 NO 的网络中的每个 UME 通过电化学沉积聚（丁香酚）和聚（苯酚）的薄层进行电化学修饰。在磷酸盐缓冲溶液（PBS，pH = 7.4）和其他适应生物细胞培养的缓冲液中，在 0.8 V 对 Ag/AgCl 进行安培检测，使用 NO 供体来检测 NO。专门用于检测 ONOO(-)的 UME 网络无需进一步表面化学修饰，未涂层的金电极在 -0.1 V 对 Ag/AgCl 下工作以检测 PBS 中 ONOOH 的还原。还检查了两种传感器对主要具有生物学相关性的干扰分析物的选择性问题。还实现了在 PBS 中同时检测 NO 和 ONOO(-)。
