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采用快速扫描循环伏安法提高多巴胺检测灵敏度。

Higher sensitivity dopamine measurements with faster-scan cyclic voltammetry.

机构信息

Department of Chemistry, University of North Carolina at Chapel Hill, 27599, United States.

出版信息

Anal Chem. 2011 May 1;83(9):3563-71. doi: 10.1021/ac200143v. Epub 2011 Apr 7.

Abstract

Fast-scan cyclic voltammetry (FSCV) with carbon-fiber microelectrodes has been successfully used to detect catecholamine release in vivo. Generally, waveforms with anodic voltage limits of 1.0 or 1.3 V (vs Ag/AgCl) are used for detection. The 1.0 V excursion provides good temporal resolution but suffers from a lack of sensitivity. The 1.3 V excursion increases sensitivity but also increases response time, which can blur the detection of neurochemical events. Here, the scan rate was increased to improve the sensitivity of the 1.0 V excursion while maintaining the rapid temporal response. However, increasing scan rate increases both the desired faradaic current response and the already large charging current associated with the voltage sweep. Analog background subtraction was used to prevent the analog-to-digital converter from saturating from the high currents generated with increasing scan rate by neutralizing some of the charging current. In vitro results with the 1.0 V waveform showed approximately a 4-fold increase in signal-to-noise ratio with maintenance of the desired faster response time by increasing scan rate up to 2400 V/s. In vivo, stable stimulated release was detected with an approximate 4-fold increase in peak current. The scan rate of the 1.3 V waveform was also increased, but the signal was unstable with time in vitro and in vivo. Adapting the 1.3 V triangular wave into a sawhorse design prevented signal decay and increased the faradaic response. The use of the 1.3 V sawhorse waveform decreased the detection limit of dopamine with FSCV to 0.96 ± 0.08 nM in vitro and showed improved performance in vivo without affecting the neuronal environment. Electron microscopy showed dopamine sensitivity is in a quasi-steady state with carbon-fiber microelectrodes scanned to potentials above 1.0 V.

摘要

快速扫描循环伏安法(FSCV)结合碳纤维微电极已成功用于检测体内儿茶酚胺的释放。通常,使用阳极电压限制为 1.0 或 1.3 V(相对于 Ag/AgCl)的波形进行检测。1.0 V 偏移量提供了良好的时间分辨率,但灵敏度不足。1.3 V 偏移量增加了灵敏度,但也增加了响应时间,这可能会使神经化学事件的检测变得模糊。在这里,扫描速率增加,以提高 1.0 V 偏移量的灵敏度,同时保持快速的时间响应。然而,增加扫描速率会增加所需的法拉第电流响应和与电压扫描相关的已经很大的充电电流。模拟背景减法用于防止模拟-数字转换器因扫描速率增加而产生的高电流而饱和,通过中和一些充电电流来实现。使用 1.0 V 波形的体外结果表明,通过将扫描速率提高到 2400 V/s,在保持所需更快响应时间的同时,信号噪声比大约增加了 4 倍。在体内,稳定的刺激释放被检测到,峰值电流大约增加了 4 倍。1.3 V 波形的扫描速率也增加了,但体外和体内的信号随时间不稳定。将 1.3 V 三角波改编成锯齿形设计防止了信号衰减,并增加了法拉第响应。使用 1.3 V 锯齿波波形降低了 FSCV 对多巴胺的检测限,在体外达到 0.96 ± 0.08 nM,并在不影响神经元环境的情况下改善了体内性能。电子显微镜显示,当碳纤维微电极扫描到 1.0 V 以上的电位时,多巴胺的灵敏度处于准稳态。

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