Wlodkowic Donald, Telford William, Skommer Joanna, Darzynkiewicz Zbigniew
BioMEMS Research Group, Department of Chemistry, University of Auckland, Auckland, New Zealand.
Methods Cell Biol. 2011;103:55-98. doi: 10.1016/B978-0-12-385493-3.00004-8.
A cell undergoing apoptosis demonstrates multitude of characteristic morphological and biochemical features, which vary depending on the inducer of apoptosis, cell type and the "time window" at which the process of apoptosis is observed. Because the gross majority of apoptotic hallmarks can be revealed by flow and image cytometry, the cytometric methods become a technology of choice in diverse studies of cellular demise. Variety of cytometric methods designed to identify apoptotic cells, detect particular events of apoptosis and probe mechanisms associated with this mode of cell death have been developed during the past two decades. In the present review, we outline commonly used methods that are based on the assessment of mitochondrial transmembrane potential, activation of caspases, DNA fragmentation, and plasma membrane alterations. We also present novel developments in the field such as the use of cyanine SYTO and TO-PRO family of probes. Strategies of selecting the optimal multiparameter approaches, as well as potential difficulties in the experimental procedures, are thoroughly summarized.
正在经历凋亡的细胞表现出众多特征性的形态和生化特征，这些特征因凋亡诱导剂、细胞类型以及观察凋亡过程的“时间窗”而异。由于绝大多数凋亡特征可通过流式细胞术和图像细胞术揭示，因此细胞计数方法成为细胞死亡各种研究中的首选技术。在过去二十年中，已开发出多种用于识别凋亡细胞、检测凋亡特定事件以及探究与这种细胞死亡模式相关机制的细胞计数方法。在本综述中，我们概述了基于评估线粒体跨膜电位、半胱天冬酶激活、DNA片段化和质膜改变的常用方法。我们还介绍了该领域的新进展，例如使用花青SYTO和TO-PRO探针家族。本文全面总结了选择最佳多参数方法的策略以及实验过程中可能遇到的困难。
