Department of Neuroscience, NIS Centre of Excellence University of Torino, Corso Raffaello 30, 10125 Torino, Italy.
Biosens Bioelectron. 2010 Sep 15;26(1):92-8. doi: 10.1016/j.bios.2010.05.017. Epub 2010 May 19.
The quantal release of oxidizable molecules can be successfully monitored by means of polarized carbon fiber microelectrodes (CFEs) positioned in close proximity to the cell membrane. To partially overcome certain CFE limitations, mainly related to their low spatial resolution and lack of optical transparency, we developed a planar boron-doped nanocrystalline diamond (NCD) prototype, grown on a transparent sapphire wafer. Responsiveness to applied catecholamines as well as the electrochemical and optical properties of the NCD-based device were first characterized by cyclic voltammetry and optical transmittance measurements. By stimulating chromaffin cells positioned on the device with external KCl, well-resolved quantal exocytotic events could be detected either from one NCD microelectrode, or simultaneously from an array of four microelectrodes, indicating that the chip is able to monitor secretory events (amperometric spikes) from a number of isolated chromaffin cells. Spikes detected by the planar NCD device had comparable amplitudes, kinetics and vesicle diameter distributions as those measured by conventional CFEs from the same chromaffin cell.
可通过将位置贴近细胞膜的极化碳纤维微电极 (CFEs) 成功监测可氧化分子的量子释放。为了部分克服某些 CFE 的局限性,主要涉及低空间分辨率和缺乏光学透明度,我们开发了一种平面掺硼纳米晶金刚石 (NCD) 原型,生长在透明蓝宝石晶片上。通过循环伏安法和光透过率测量,首先对基于 NCD 的设备的应用儿茶酚胺响应以及电化学和光学性质进行了表征。通过用外部 KCl 刺激置于设备上的嗜铬细胞,可从一个 NCD 微电极或同时从四个微电极阵列中检测到分辨率良好的量子胞吐事件,表明该芯片能够监测来自多个分离的嗜铬细胞的分泌事件 (安培峰)。平面 NCD 设备检测到的尖峰与从同一嗜铬细胞的常规 CFE 测量的尖峰具有可比的幅度、动力学和囊泡直径分布。