Apoptosis and beyond: cytometry in studies of programmed cell death.
Apoptosis and beyond: cytometry in studies of programmed cell death.
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DOI：
10.1016/b978-0-12-385493-3.00004-8
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发表时间：
2011
影响因子：
--
通讯作者：
Darzynkiewicz, Zbigniew
中科院分区：
文献类型：
--
作者：
Wlodkowic, Donald;Telford, William;Skommer, Joanna;Darzynkiewicz, Zbigniew
经历细胞凋亡的细胞表现出多种特征性形态和生化特征，这些特征根据细胞凋亡诱导剂、细胞类型和观察细胞凋亡过程的“时间窗口”而变化。由于绝大多数细胞凋亡标志可以通过流式细胞术和图像细胞术来揭示，因此细胞计数方法成为各种细胞死亡研究的首选技术。在过去的二十年中，已经开发了多种旨在识别凋亡细胞、检测特定的凋亡事件以及探测与这种细胞死亡模式相关的机制的细胞计数方法。在本综述中，我们概述了基于线粒体跨膜电位、半胱天冬酶激活、DNA 片段化和质膜改变评估的常用方法。我们还介绍了该领域的新进展，例如菁 SYTO 和 TO-PRO 系列探针的使用。彻底总结了选择最佳多参数方法的策略以及实验过程中的潜在困难。
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.