Matsushita T, Brendzel A M, Shotola M A, Groh K R
Biophys J. 1982 Jul;39(1):41-7. doi: 10.1016/S0006-3495(82)84488-3.
Suspension of mouse myeloma cells in phosphate buffered saline (PBS) induced a significant amount of cell death. The lethal effects of PBS include an increase in cell lysis, a decreased ability of cells to exclude trypan blue, and a decrease in the colony-forming ability of these cells. Dead cells were also detected on a Coulter counter by the increase in the fraction of cells with a smaller electrical size distribution (ESD). Comparing mixtures of live and dead cells by ESD and trypan-blue exclusion showed a high correlation of electrical size with viability (correlation coefficient = 0.98). Sizing of PBS-treated cells by light microscopy suggested that the altered ESD of the PBS-treated cells was due to a downward shift in the volume distribution. Light-scattering experiments also suggested a decrease in the size of cells after PBS treatment. An increase in permeability of the cell membrane may also contribute to these results. We conclude that electrical sizing is an excellent indicator of physical changes that result from PBS-induced cell death, and is an effective method for distinguishing live and dead mouse myeloma cells after PBS treatment.
将小鼠骨髓瘤细胞悬浮于磷酸盐缓冲盐水（PBS）中会导致大量细胞死亡。PBS的致死作用包括细胞裂解增加、细胞排斥台盼蓝的能力下降以及这些细胞集落形成能力降低。通过库尔特计数器检测到，电尺寸分布（ESD）较小的细胞比例增加，表明存在死细胞。通过ESD和台盼蓝排斥法比较活细胞和死细胞混合物，结果显示电尺寸与活力高度相关（相关系数 = 0.98）。光学显微镜观察经PBS处理的细胞大小，结果表明经PBS处理的细胞ESD改变是由于体积分布向下偏移所致。光散射实验也表明PBS处理后细胞尺寸减小。细胞膜通透性增加也可能导致这些结果。我们得出结论，电尺寸分析是PBS诱导细胞死亡所导致物理变化的极佳指标，并且是区分PBS处理后活的和死的小鼠骨髓瘤细胞的有效方法。
