Centre for Biochemical Engineering, School of Chemical Engineering, University of Birmingham, Edgbaston, Birmingham, B 15 2 TT, UK.
Cytotechnology. 1997 Jul;24(2):161-8. doi: 10.1023/A:1007910920355.
A simple, rapid and reliable method has been developed for assessing the number and viability of cells, as well as cell size, in suspension culture by the use of flow cytometry. Propidium iodide exclusion is used for viability determination and fluorescent beads serve as an internal standard for cell enumeration. The main advantages of this method are its ability to handle a large number of samples with a high degree of precision and its specificity in detecting viable cells quantitatively in a heterogeneous culture of living and dead cells and debris. The method shows only a fraction of the variation found in the haemacytometer/trypan blue counting method due to its very low operator dependence. CHO - Chinese hamster ovary; FCS - Foetal calf serum; FS - Forward scatter light; MTT - 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide; NCS - newborn calf serum; PBS - Phosphate buffered saline; PI - Propidium iodide; SS - Side scatter light.
已开发出一种简单、快速和可靠的方法，通过使用流式细胞术评估悬浮培养物中的细胞数量和活力以及细胞大小。碘化丙啶排除用于测定活力，荧光珠用作细胞计数的内部标准。该方法的主要优点是能够高精度处理大量样品，并且能够特异性地定量检测活细胞在活细胞和死细胞以及碎片的异质培养物中的存在。与由于操作人员依赖性低而导致的相差显微镜/台盼蓝计数法相比，该方法仅显示出一小部分变化。CHO-中国仓鼠卵巢；FCS-胎牛血清；FS-前向散射光；MTT-3-(4,5-二甲基噻唑基-2)-2,5-二苯基四氮唑溴盐；NCS-新生牛血清；PBS-磷酸盐缓冲盐水；PI-碘化丙啶；SS-侧向散射光。
