Department of Micro- and Nanotechnology, Technical University of Denmark, Ørsted Plads 345E, 2800 Kgs. Lyngby, Denmark.
Analyst. 2013 Jul 7;138(13):3651-9. doi: 10.1039/c3an00085k.
A surface modification of interdigitated gold microelectrodes (IDEs) with a doped polypyrrole (PPy) film for detection of dopamine released from populations of differentiated PC12 cells is presented. A thin PPy layer was potentiostatically electropolymerized from an aqueous pyrrole solution onto electrode surfaces. The conducting polymer film was doped during electropolymerization by introducing counter-ions in the monomer solution. Several counter-ions were tested and the resulting electrode modifications were characterized electrochemically to find the optimal dopant that increases sensitivity in dopamine detection. Overoxidation of the PPy films was shown to contribute to a significant enhancement in sensitivity to dopamine. The changes caused by overoxidation in the electrochemical behavior and electrode morphology were investigated using cyclic voltammetry and SEM as well as AFM, respectively. The optimal dopant for dopamine detection was found to be polystyrene sulfonate anion (PSS(-)). Rat pheochromocytoma (PC12) cells, a suitable model to study exocytotic dopamine release, were differentiated on IDEs functionalized with an overoxidized PSS(-)-doped PPy film. The modified electrodes were used to amperometrically detect dopamine released by populations of cells upon triggering cellular exocytosis with an elevated K(+) concentration. A comparison between the generated current on bare gold electrodes and gold electrodes modified with overoxidized doped PPy illustrates the clear advantage of the modification, yielding 2.6-fold signal amplification. The results also illustrate how to use cell population based dopamine exocytosis measurements to obtain biologically significant information that can be relevant in, for instance, the study of neural stem cell differentiation into dopaminergic neurons.
一种用于检测分化的 PC12 细胞群体释放多巴胺的叉指金微电极(IDE)表面修饰方法,该方法通过掺杂聚吡咯(PPy)薄膜实现。通过在电极表面上从水性吡咯溶液中电化学聚合,制备了一层薄的 PPy 层。在聚合过程中,通过在单体溶液中引入抗衡离子,对导电聚合物薄膜进行掺杂。测试了几种抗衡离子,并通过电化学方法对所得电极修饰进行了表征,以找到增加多巴胺检测灵敏度的最佳掺杂剂。研究表明,PPy 薄膜的过度氧化有助于显著提高对多巴胺的灵敏度。使用循环伏安法和 SEM 以及 AFM 分别研究了电化学行为和电极形貌的过度氧化引起的变化。发现聚苯乙烯磺酸盐阴离子(PSS(-))是多巴胺检测的最佳掺杂剂。嗜铬细胞瘤(PC12)细胞是研究胞吐释放多巴胺的合适模型,将其分化在功能化的过氧化掺杂 PSS(-) - PPy 薄膜上的 IDE 上。使用修饰后的电极通过升高的 K(+)浓度触发细胞胞吐作用,对细胞群体释放的多巴胺进行安培检测。裸金电极和过氧化掺杂 PPy 修饰的金电极上产生的电流的比较说明了修饰的明显优势,产生了 2.6 倍的信号放大。该结果还说明了如何使用基于细胞群体的多巴胺胞吐作用测量来获得生物学上有意义的信息,这些信息可能与神经干细胞分化为多巴胺能神经元的研究等有关。