Belloc F, Dumain P, Boisseau M R, Jalloustre C, Reiffers J, Bernard P, Lacombe F
Laboratoire d'Hématologie, Hôpital Haut Lévêque, Pessac, France.
Cytometry. 1994 Sep 1;17(1):59-65. doi: 10.1002/cyto.990170108.
A flow cytometric method to detect apoptotic cells is described. This method is based on the detection of differences in chromatin condensation with Hoechst 33342 as a probe and the detection of dead cells with propidium iodide as a probe for membrane damage. By this method it was possible to detect, in the same sample and at the same time, intact cells, cells undergoing apoptosis, and dead cells resulting from apoptotic and/or necrotic processes. The method was successfully applied to the detection of apoptotic cells in two human cell models: cultured polymorphonuclear cells and the U937 cell line treated with antitumoral drugs. Staining specificity for apoptotic cells was controlled by cell sorting of the presumed apoptotic population, followed by morphologic examination or DNA analysis of the sorted populations. The usefulness of such a method is discussed in terms of applications in the analysis of heterogeneous clinical samples, populations with low DNA degradation during apoptosis, and cell cycle position of the apoptotic cells.
本文描述了一种用于检测凋亡细胞的流式细胞术方法。该方法基于以Hoechst 33342为探针检测染色质凝聚差异，以及以碘化丙啶为探针检测膜损伤来识别死细胞。通过这种方法，可以在同一样本中同时检测完整细胞、正在经历凋亡的细胞以及由凋亡和/或坏死过程导致的死细胞。该方法已成功应用于两种人类细胞模型中凋亡细胞的检测：培养的多形核细胞和用抗肿瘤药物处理的U937细胞系。通过对假定的凋亡群体进行细胞分选，随后对分选群体进行形态学检查或DNA分析，来控制对凋亡细胞的染色特异性。从该方法在异质性临床样本分析、凋亡过程中DNA降解程度低的群体以及凋亡细胞的细胞周期位置分析中的应用等方面，讨论了这种方法的实用性。
