Improved Surface-Patterned Platinum Microelectrodes for the Study of Exocytotic Events
Improved Surface-Patterned Platinum Microelectrodes for the Study of Exocytotic Events
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DOI：
10.1021/ac900674g
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发表时间：
2009-11-01
影响因子：
7.4
通讯作者：
Lindau, Manfred
中科院分区：
文献类型：
--
作者：
Berberian, Khajak;Kisler, Kassandra;Lindau, Manfred
采用接触光刻技术制备了300 nm厚熔融二氧化硅绝缘的表面图案化铂微电极。这些电极具有低噪声，可用于用恒电位安培法和快速扫描循环伏安法监测嗜铬细胞的单个囊泡胞吐。电流尖峰参数与常规碳纤维电极的结果一致。用铂电极采集的儿茶酚胺伏安图显示氧化还原峰，半峰宽接近45 mV，比碳纤维电极记录的结果更锐利。对于铂电极和碳纤维电极，伏安法测量的释放事件的时间过程相似。熔融二氧化硅绝缘的铂电极可以反复清洗和重复使用，并允许掺入微米级精密表面图案化的聚D-赖氨酸。使用多聚D-赖氨酸功能化装置刺激肥大细胞并记录单次释放事件，而不预先加载5-羟色胺。因此，微细加工的铂电极能够以高分辨率记录单个胞吐事件，并且应该适合于高度平行的电极阵列，从而允许同时测量来自多个电池的单个事件。
Surface-patterned platinum microelectrodes insulated with 300 nm thick fused silica were fabricated using contact photolithography. These electrodes exhibit low noise and were used for monitoring single vesicle exocytosis from chromaffin cells by constant potential amperometry as well as fast-scan cyclic voltammetry. Amperometric spike parameters were consistent with those obtained with conventional carbon fiber electrodes. Catecholamine voltammograms acquired with platinum electrodes exhibited redox peaks with full width at half-maximum of similar to 45 mV, much sharper than those of carbon fiber electrode recordings. The time course of voltammetrically measured release events was similar for platinum and carbon fiber electrodes. The fused-silica-insulated platinum electrodes could be cleaned and reused repetitively and allowed incorporation of micrometer precision surface-patterned poly-D-lysine. Poly-D-lysine-functionalized devices were applied to stimulate mast cells and record single release events without serotonin pre-loading. Microfabricated platinum electrodes are thus able to record single exocytotic events with high resolution and should be suitable for highly parallel electrode arrays allowing simultaneous measurements of single events from multiple cells.