Dankberg F, Persidsky M D
Cryobiology. 1976 Aug;13(4):430-2. doi: 10.1016/0011-2240(76)90098-5.
An assay of granulocyte viability has been developed which yields information about rwo important cell parameters, cell membrane integrity and phagocytic activity. The assay is based on the fact that only live cells can accumulate fluorescein, which is enzymatically split from the nonfluorescent substrate fluorescein diacetate. Dead cells, on the other hand, become permeable to the fluorescent red dye ethidium bromide. When cells are exposed first to opsonized zymosan particles, which they can phagocytize, then to a combination of these fluorescent dyes, one can distinguish microscopically between dead cells with fluorescent red nuclei, live cells which fluoresce green, and live cells with phagocytic function which are swollen with the pink zymosan particles in a green fluorescing cytoplasm. This assay takes 20--30 min and can be used to distinguish different degrees of cellular damage after cryopreservation.
已开发出一种粒细胞活力测定法，该方法可提供有关两个重要细胞参数的信息，即细胞膜完整性和吞噬活性。该测定法基于这样一个事实：只有活细胞才能积累荧光素，荧光素是从无荧光的底物荧光素二乙酸酯经酶解而来。另一方面，死细胞对红色荧光染料溴化乙锭具有通透性。当细胞首先暴露于它们可以吞噬的调理酵母聚糖颗粒，然后再暴露于这些荧光染料的组合时，在显微镜下可以区分出细胞核呈红色荧光的死细胞、发绿色荧光的活细胞以及具有吞噬功能且在绿色荧光细胞质中充满粉红色酵母聚糖颗粒而肿胀的活细胞。该测定需要20 - 30分钟，可用于区分冷冻保存后不同程度的细胞损伤。
