Dengler Adam K, Wightman R Mark, McCarty Gregory S
Department of Biomedical Engineering, North Carolina State University , Raleigh, North Carolina 27695, United States.
Department of Chemistry, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina 27599, United States.
Anal Chem. 2015 Oct 20;87(20):10556-64. doi: 10.1021/acs.analchem.5b02866. Epub 2015 Sep 29.
Fast-scan cyclic voltammetry (FSCV) has attracted attention for studying in vivo neurotransmission due to its subsecond temporal resolution, selectivity, and sensitivity. Traditional FSCV measurements use background subtraction to isolate changes in the local electrochemical environment, providing detailed information on fluctuations in the concentration of electroactive species. This background subtraction removes information about constant or slowly changing concentrations. However, determination of background concentrations is still important for understanding functioning brain tissue. For example, neural activity is known to consume oxygen and produce carbon dioxide which affects local levels of oxygen and pH. Here, we present a microfabricated microelectrode array which uses FSCV to detect the absolute levels of oxygen and pH in vitro. The sensor is a collector-generator electrode array with carbon microelectrodes spaced 5 μm apart. In this work, a periodic potential step is applied at the generator producing transient local changes in the electrochemical environment. The collector electrode continuously performs FSCV enabling these induced changes in concentration to be recorded with the sensitivity and selectivity of FSCV. A negative potential step applied at the generator produces a transient local pH shift at the collector. The generator-induced pH signal is detected using FSCV at the collector and correlated to absolute solution pH by postcalibration of the anodic peak position. In addition, in oxygenated solutions a negative potential step at the generator produces hydrogen peroxide by reducing oxygen. Hydrogen peroxide is detected with FSCV at the collector electrode, and the magnitude of the oxidative peak is proportional to absolute oxygen concentrations. Oxygen interference on the pH signal is minimal and can be accounted for with a postcalibration.
快速扫描循环伏安法(FSCV)因其亚秒级的时间分辨率、选择性和灵敏度,在研究体内神经传递方面备受关注。传统的FSCV测量使用背景扣除来分离局部电化学环境的变化,提供有关电活性物质浓度波动的详细信息。这种背景扣除消除了关于恒定或缓慢变化浓度的信息。然而,背景浓度的测定对于理解脑组织的功能仍然很重要。例如,已知神经活动会消耗氧气并产生二氧化碳,这会影响局部氧气水平和pH值。在此,我们展示了一种微制造的微电极阵列,它使用FSCV在体外检测氧气和pH的绝对水平。该传感器是一个集电极 - 发生器电极阵列,碳微电极间距为5μm。在这项工作中,在发生器处施加周期性电势阶跃,在电化学环境中产生瞬态局部变化。集电极连续执行FSCV,从而能够以FSCV的灵敏度和选择性记录这些诱导的浓度变化。在发生器处施加的负电势阶跃会在集电极处产生瞬态局部pH偏移。利用集电极处的FSCV检测发生器诱导的pH信号,并通过阳极峰位置的后校准将其与绝对溶液pH相关联。此外,在含氧溶液中,发生器处的负电势阶跃通过还原氧气产生过氧化氢。在集电极处用FSCV检测过氧化氢,氧化峰的大小与绝对氧气浓度成正比。氧气对pH信号的干扰最小,可以通过后校准来解释。