Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, Ontario M5S 3H6, Canada.
Analyst. 2010 Feb;135(2):289-95. doi: 10.1039/b919430b. Epub 2009 Dec 21.
Isolation of neurons from animal tissue is an important aspect of understanding basic biochemical processes such as the action of hormones and neurotransmitters. In the present work, the focus is on an effort to evaluate the utility of acoustic wave physics for the study of such cells. Immortalised hypothalamic neuronal cells from mouse embryos were cultured on the surface of the gold electrode of a 9.0 MHz thickness-shear mode acoustic wave sensor. These cells, which are clonal, are imposed on the surface of the device at a confluence in the range of 80-100%. The coated sensor is incorporated into a flow-injection configuration such that electrolytes can be introduced in order to examine their effects through measurement by network analysis. Both series resonance frequency, fs, and motional resistance, R(m), were measured in a number of experiments involving the injection of KCl and NaCl into the sensor-neuron system. The various responses to these electrolytes were interpreted in terms of changes in cellular structure associated with the depolarization process. The sensor-neuron system was found to elicit different responses to the addition of KCl and NaCl. Preliminary findings indicate that the TSM sensor does not purely measure changes in the membrane potential upon KCl addition. Typical changes in fs for 15 mM, 30 mM and 60 mM KCl additions were 54 +/- 15, 80 +/- 26 and 142 +/- 58 Hz (mean +/- standard deviation) respectively. Typical changes in R(m) for these KCl additions were 7 +/- 3, 13 +/- 4 and 23 +/- 6 Omega, respectively. These results were concluded after 17 runs at each concentration. Despite the large relative standard deviations, the dependence of f(s) and R(m) with respect to concentration was apparent. Controls performed by coating the TSM sensor with laminin or a cell attachment matrix showed no significant changes in either f(s) or R(m) for the same solutions tested on the sensor-neuron system.
从动物组织中分离神经元是理解基本生化过程（如激素和神经递质的作用）的重要方面。在本工作中，重点是评估声波物理在研究此类细胞中的应用。从鼠胚中分离出的永生下丘脑神经元细胞被培养在 9.0MHz 厚度剪切模式声波传感器的金电极表面上。这些细胞是克隆的，在设备表面以 80-100%的汇合度被施加。涂覆的传感器被纳入流动注射配置中，以便通过网络分析测量来检查电解质的影响。在涉及将 KCl 和 NaCl 注入传感器-神经元系统的多次实验中，测量了串联共振频率 fs 和运动电阻 R(m)。根据与去极化过程相关的细胞结构变化，解释了对这些电解质的各种响应。发现传感器-神经元系统对添加 KCl 和 NaCl 的反应不同。初步研究结果表明，TSM 传感器并不仅仅测量 KCl 添加后膜电位的变化。对于 15mM、30mM 和 60mM KCl 添加，fs 的典型变化分别为 54 +/- 15、80 +/- 26 和 142 +/- 58Hz（平均值 +/- 标准偏差）。对于这些 KCl 添加，Rm 的典型变化分别为 7 +/- 3、13 +/- 4 和 23 +/- 6Omega。在每个浓度下进行了 17 次运行后得出了这些结果。尽管相对标准偏差较大，但 fs 和 R(m) 与浓度的依赖性是明显的。通过用层粘连蛋白或细胞附着基质涂覆 TSM 传感器进行的对照实验表明，对于在传感器-神经元系统上测试的相同溶液，fs 或 R(m) 没有明显变化。
