Pancrazio Joseph J, Gray Samuel A, Shubin Yura S, Kulagina Nadezhda, Cuttino David S, Shaffer Kara M, Eisemann Kevin, Curran Anthony, Zim Bret, Gross Guenter W, O'Shaughnessy Thomas J
Center for Bio/Molecular Science and Engineering, Code 6900, Naval Research Laboratory, Washington, DC 20375, USA.
Biosens Bioelectron. 2003 Oct 1;18(11):1339-47. doi: 10.1016/s0956-5663(03)00092-7.
Cultured neuronal networks, which have the capacity to respond to a wide range of neuroactive compounds, have been suggested to be useful for both screening known analytes and unknown compounds for acute neuropharmacologic effects. Extracellular recording from cultured neuronal networks provides a means for extracting physiologically relevant activity, i.e. action potential firing, in a noninvasive manner conducive for long-term measurements. Previous work from our laboratory described prototype portable systems capable of high signal-to-noise extracellular recordings from cardiac myocytes. The present work describes a portable system tailored to monitoring neuronal extracellular potentials that readily incorporates standardized microelectrode arrays developed by and in use at the University of North Texas. This system utilizes low noise amplifier and filter boards, a two-stage thermal control system with integrated fluidics and a graphical user interface for data acquisition and control implemented on a personal computer. Wherever possible, off-the-shelf components have been utilized for system design and fabrication. During use with cultured neuronal networks, the system typically exhibits input referred noise levels of only 4-6 microVRMS, such that extracellular potentials exceeding 40 microV can be readily resolved. A flow rate of up to 1 ml/min was achieved while the cell recording chamber temperature was maintained within a range of 36-37 degrees C. To demonstrate the capability of this system to resolve small extracellular potentials, pharmacological experiments with cultured neuronal networks have been performed using ion channel blockers, tetrodotoxin and tityustoxin. The implications of the experiments for neurotoxin detection are discussed.
培养的神经网络能够对多种神经活性化合物作出反应,已被认为可用于筛选已知分析物和未知化合物的急性神经药理作用。从培养的神经网络进行细胞外记录提供了一种以无创方式提取生理相关活动(即动作电位发放)的手段,有利于进行长期测量。我们实验室之前的工作描述了能够对心肌细胞进行高信噪比细胞外记录的原型便携式系统。本工作描述了一种专门用于监测神经元细胞外电位的便携式系统,该系统很容易集成了由北德克萨斯大学开发并正在使用的标准化微电极阵列。该系统利用低噪声放大器和滤波器板、带有集成流体ics的两级热控制系统以及在个人计算机上实现的数据采集和控制的图形用户界面。在可能的情况下,系统设计和制造采用了现成的组件。在与培养的神经网络一起使用时,该系统通常表现出仅4 - 6微伏均方根值的输入参考噪声水平,使得超过40微伏的细胞外电位能够很容易地分辨出来。在细胞记录室温度保持在36 - 37摄氏度范围内的情况下,实现了高达1毫升/分钟的流速。为了证明该系统分辨小细胞外电位的能力,使用离子通道阻滞剂、河豚毒素和墨西哥蝎子毒素对培养的神经网络进行了药理学实验。讨论了这些实验对神经毒素检测的意义。