Behavioural Neuroscience Laboratory, Department of Experimental Medical Sciences, Lund University, Lund, Sweden.
Methods Mol Biol. 2021;2352:201-226. doi: 10.1007/978-1-0716-1601-7_14.
The detection of neurotransmitter release from reprogrammed human cell is an important demonstration of their functionality. Electrochemistry has the distinct advantages over alternative methods that it allows for the measuring of the analyte of interest at a high temporal resolution. This is necessary for fast events, such as neurotransmitter release and reuptake, which happen in the order of milliseconds to seconds. The precise description of these kinetic events can lead to insights into the function of cells in health and disease and allows for the exploration of events that might be missed using methods that look at absolute concentration values or methods that have a slower sampling rate. In the present chapter, we describe the use of constant potential amperometry and enzyme-coated multielectrode arrays for the detection of glutamate in vitro. These biosensors have the distinct advantage of "self-referencing," a method providing high selectivity while retaining outstanding temporal resolution. Here, we provide a step-by-step user guide for a commercially available system and its application for in vitro systems such as reprogrammed cells.
检测重编程人类细胞中的神经递质释放是证明其功能的重要手段。与其他方法相比,电化学具有明显的优势,它可以以高时间分辨率测量感兴趣的分析物。这对于快速事件(如神经递质的释放和再摄取)是必要的,这些事件发生在毫秒到秒的时间范围内。对这些动力学事件的精确描述可以深入了解健康和疾病状态下细胞的功能,并允许探索使用绝对浓度值方法或采样率较慢的方法可能会错过的事件。在本章中,我们描述了使用恒电位安培法和酶涂覆的多电极阵列检测体外谷氨酸。这些生物传感器具有“自我参考”的明显优势,这种方法提供了高选择性,同时保持出色的时间分辨率。在这里,我们提供了一个商业可用系统的分步用户指南及其在体外系统(如重编程细胞)中的应用。