Frankfurt O S
Exp Cell Res. 1983 Apr 1;144(2):478-82. doi: 10.1016/0014-4827(83)90428-7.
A new method was developed for selective measurement of DNA distributions in viable cell populations. The method is based on the fact that non-viable cells lose membrane integrity and treatment of such cells with DNase should remove their DNA. The DNase-treated cells were stained with DNA fluorochrome 4'-6-diamidino-2-phenylindole (DAPI) in the presence of Triton X-100. DNA distribution was measured by flow cytometry prior to and after treatment with DNase. Percentage of cells stained after DNase treatment was considered as an index of cell viability. Optimal conditions for DNase treatment and application of DNase exclusion test for the analysis of spontaneous cell death, selective death of cells arrested in S/G2 phases, instant cell disintegration induced by cytotoxic compounds and cell death induced by hyperthermia are described.
开发了一种用于选择性测量活细胞群体中DNA分布的新方法。该方法基于以下事实：非活细胞会丧失膜完整性，用脱氧核糖核酸酶（DNase）处理此类细胞应能去除其DNA。在Triton X-100存在的情况下，用DNA荧光染料4′-6-二脒基-2-苯基吲哚（DAPI）对经DNase处理的细胞进行染色。在用DNase处理之前和之后，通过流式细胞术测量DNA分布。DNase处理后染色的细胞百分比被视为细胞活力指标。描述了DNase处理的最佳条件以及将DNase排除试验应用于分析自发细胞死亡、停滞在S/G2期细胞的选择性死亡、细胞毒性化合物诱导的即时细胞解体和热疗诱导的细胞死亡。
