Zimmermann U, Schulz J, Pilwat G
Biophys J. 1973 Oct;13(10):1005-13. doi: 10.1016/S0006-3495(73)86041-2.
Dielectric breakdown of cell membranes and, in response, transcellular ion flows were measured in Escherichia coli B 163 and B 525 using a Coulter counter as the detector with a hydrodynamic jet focusing close to the orifice of the counter. Plotting the relative pulse height for compensated amplification of a certain size of the cells against increasing detector current, a rather sharp bend within the linear function was found, which did not occur when measuring fixed cells or polystyrene latex. The start current for transcellular ion flow causing the change of the slope is different for the potassium-deficient mutant B 525 in comparison with the wild-type B 163, indicating a change in the membrane structure of B 525 by mutation and demonstrating the sensitivity of the method for studying slight changes in membrane structure in general. The theoretical size distributions for two current values in the range of transcellular ion flow were constructed from the true size distribution at low detector currents, assuming an idealized sharp changeover of the bacterial conductivity from zero to one-third of the electrolyte conductivity.
使用库尔特计数器作为检测器,在靠近计数器孔口处采用流体动力喷射聚焦,测量了大肠杆菌B 163和B 525细胞膜的介电击穿以及相应的跨细胞离子流。将特定大小细胞经补偿放大后的相对脉冲高度相对于增加的检测器电流作图,发现在线性函数内有一个相当明显的弯曲,而在测量固定细胞或聚苯乙烯胶乳时并未出现这种情况。与野生型B 163相比,缺钾突变体B 525中引起斜率变化的跨细胞离子流起始电流不同,这表明B 525的膜结构因突变而发生了变化,总体上证明了该方法对研究膜结构细微变化的敏感性。在跨细胞离子流范围内,根据低检测器电流下的真实大小分布,假设细菌电导率从零到电解质电导率的三分之一有一个理想化的急剧转变,构建了两个电流值的理论大小分布。