Cahill P S, Walker Q D, Finnegan J M, Mickelson G E, Travis E R, Wightman R M
Department of Chemistry, University of North Carolina, Chapel Hill 27599-3290, USA.
Anal Chem. 1996 Sep 15;68(18):3180-6. doi: 10.1021/ac960347d.
Many of the molecules involved in biological signaling processes are easily oxidized and have been monitored by electrochemical methods. Temporal response, spatial considerations, and sensitivity of the electrodes must be optimized for the specific biological application. To monitor exocytosis from single cells in culture, constant potential amperometry offers the best temporal resolution, and a low-noise picoammeter improves the detection limits. Smaller electrodes, with 1-micron diameters, provided spatial resolution sufficient to identify the locations of release sites on the surface of single cells. For the study of neurotransmitter release in vivo, larger cylindrical microelectrodes are advantageous because the secreted molecules come from multiple terminals near the electrode, and the greater amounts lead to a larger signal that emerges from the Johnson noise of the current amplifier. With this approach, dopamine release elicited by two electrical stimulus pulses at 10 Hz was detected with fastscan cyclic voltammetry in vivo. Nafion-coated elliptical electrodes have previously been shown to be incapable of detecting such concentration changes without extensive signal averaging. In addition, we demonstrate that high-pass filtering (200 Hz) of cyclic voltammograms recorded at 300 V/s decreases the background current and digitization noise at these microelectrodes, leading to an improved signal. Also, high-pass filtering discriminated against ascorbic acid, DOPAC, and acidic pH changes, three common interferences in vivo.
许多参与生物信号传导过程的分子很容易被氧化,并且已经通过电化学方法进行监测。对于特定的生物学应用,必须优化电极的时间响应、空间因素和灵敏度。为了监测培养的单细胞的胞吐作用,恒电位安培法提供了最佳的时间分辨率,而低噪声皮安计则提高了检测限。直径为1微米的较小电极提供了足够的空间分辨率,以识别单细胞表面释放位点的位置。对于体内神经递质释放的研究,较大的圆柱形微电极具有优势,因为分泌的分子来自电极附近的多个终端,并且量更大导致从电流放大器的约翰逊噪声中出现更大的信号。通过这种方法,在体内用快速扫描循环伏安法检测了10Hz的两个电刺激脉冲引起的多巴胺释放。以前已经证明,在没有广泛信号平均的情况下,涂有Nafion的椭圆形电极无法检测到这种浓度变化。此外,我们证明,在300V/s下记录的循环伏安图的高通滤波(200Hz)会降低这些微电极处的背景电流和数字化噪声,从而改善信号。此外,高通滤波可以区分抗坏血酸、3,4-二羟基苯乙酸和酸性pH变化,这是体内三种常见的干扰因素。