Telford William G, Komoriya Akira, Packard Beverly Z, Bagwell C Bruce
FACS Core Facilty, Experimental Transplantation and Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2011;699:203-27. doi: 10.1007/978-1-61737-950-5_10.
Flow cytometry is the most widely used technology for analyzing apoptosis. The multiparametric nature of flow cytometry allows several apoptotic characteristics to be combined in a single sample, making it a powerful tool for analyzing the complex progression of apoptotic death. This chapter provides guidelines for combining caspase detection, annexin V binding, DNA dye exclusion, and other single apoptotic assays into multiparametric assays.This approach to analyzing apoptosis provides far more information than single parameter assays that provide only an ambiguous "percent apoptotic" result, given that multiple early, intermediate and late apoptotic stages can be visualized simultaneously. This multiparametric approach is also amenable to a variety of flow cytometric instrumentation, both old and new.
流式细胞术是分析细胞凋亡最广泛使用的技术。流式细胞术的多参数特性允许在单个样本中组合多种凋亡特征,使其成为分析凋亡死亡复杂进程的强大工具。本章提供了将半胱天冬酶检测、膜联蛋白V结合、DNA染料排斥和其他单一凋亡检测组合成多参数检测的指南。这种分析细胞凋亡的方法提供的信息远比单一参数检测多,单一参数检测只能给出一个模糊的“凋亡百分比”结果,因为多个早期、中期和晚期凋亡阶段可以同时可视化。这种多参数方法也适用于各种新旧流式细胞仪。