Telford William, Tamul Karen, Bradford Jolene
Experimental Transplantation and Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland.
EMD Millipore, Inc, Billerica, Massachusetts.
Curr Protoc Cytom. 2016 Jul 1;77:9.49.1-9.49.28. doi: 10.1002/cpcy.1.
Apoptosis is an important mechanism in cell biology, playing a critical regulatory role in virtually every organ system. It has been particularly well characterized in the immune system, with roles ranging from immature immune cell development and selection to down-regulation of the mature immune response. Apoptosis is also the primary mechanism of action of anti-cancer drugs. Flow cytometry has been the method of choice for analyzing apoptosis in suspension cells for more than 25 years. Numerous assays have been devised to measure both the earliest and latest steps in the apoptotic process, from the earliest signal-transduction events to the late morphological changes in cell shape and granularity, proteolysis, and chromatin condensation. These assays are particularly powerful when combined into multicolor assays determining several apoptotic characteristics simultaneously. The multiparametric nature of flow cytometry makes this technology particularly suited to measuring apoptosis. In this unit, we will describe the four main techniques for analyzing caspase activity in apoptotic cells, combined with annexin V and cell permeability analysis. These relatively simple multiparametric assays are powerful techniques for assessing cell death. © 2016 by John Wiley & Sons, Inc.
细胞凋亡是细胞生物学中的一种重要机制，几乎在每个器官系统中都发挥着关键的调节作用。它在免疫系统中得到了特别充分的研究，其作用涵盖从未成熟免疫细胞的发育和选择到成熟免疫反应的下调。细胞凋亡也是抗癌药物的主要作用机制。25多年来，流式细胞术一直是分析悬浮细胞凋亡的首选方法。人们设计了许多检测方法来测量细胞凋亡过程中最早和最晚的步骤，从最早的信号转导事件到细胞形状、颗粒度、蛋白水解和染色质浓缩等晚期形态变化。当这些检测方法结合成多色检测同时确定几种凋亡特征时，它们特别强大。流式细胞术的多参数性质使这项技术特别适合于测量细胞凋亡。在本单元中，我们将描述分析凋亡细胞中半胱天冬酶活性的四种主要技术，并结合膜联蛋白V和细胞通透性分析。这些相对简单的多参数检测是评估细胞死亡的强大技术。© 2016约翰威立父子公司版权所有
