Dittert I, Vlachová V, Knotková H, Vitásková Z, Vyklicky L, Kress M, Reeh P W
Institute of Physiology, Prague, Czech Republic.
J Neurosci Methods. 1998 Aug 1;82(2):195-201. doi: 10.1016/s0165-0270(98)00051-x.
A technique is described that allows the application of fast temperature changes (time constant approximately 300 ms) of solutions superfusing cultured neurones under whole-cell mode of membrane current recording. Its principle is in heating the common outlet of the manifold which consists of 12 tubes connected to barrels containing test solutions of different composition. The outlet is made from a glass capillary (25 mm length, 620/350 microns outer/inner diameter) coated on the outside wall with platinum for a length of 12 mm. The heating element, a platinum layer, is electrically connected to the probe fixed to the micromanipulator used for positioning the manifold. The solutions, driven by gravity, are applied by opening electronic valves controlled either manually or in programmed sequences. The DC current for heating is controlled either manually or by external voltage command. The advantage of the technique is that the same temperature pattern can be applied to 12 different solutions. The technique is used for classifying sensory neurones in culture with respect to their sensitivity to heat and algogens; however, it is applicable to any study of the effects of increased temperature on the activity of ion channels in cultured cells.
本文描述了一种技术，该技术可在全细胞膜电流记录模式下，对灌流培养神经元的溶液施加快速温度变化（时间常数约300毫秒）。其原理是加热歧管的公共出口，该歧管由12根管子组成，这些管子连接到装有不同成分测试溶液的桶上。出口由玻璃毛细管制成（长度25毫米，外径/内径为620/350微米），外壁涂有12毫米长的铂。加热元件为铂层，与固定在用于定位歧管的微操纵器上的探头电连接。溶液在重力作用下，通过打开手动或按程序控制的电子阀来施加。加热用的直流电流可手动控制或由外部电压指令控制。该技术的优点是相同的温度模式可应用于12种不同的溶液。该技术用于根据培养的感觉神经元对热和致痛物质的敏感性进行分类；然而，它适用于任何关于温度升高对培养细胞中离子通道活性影响的研究。
