流式细胞术检测和表征细胞凋亡

Q1 Health Professions Current Protocols in Cytometry Pub Date : 2016-07-01 DOI:10.1002/cpcy.1
William Telford, Karen Tamul, Jolene Bradford
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引用次数: 13

摘要

细胞凋亡是细胞生物学中的一个重要机制,在几乎所有器官系统中都起着重要的调节作用。在免疫系统中,它的作用范围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年来,流式细胞术一直是分析悬浮细胞凋亡的首选方法。从最早的信号转导事件到细胞形状和粒度、蛋白质水解和染色质凝聚等后期形态学变化,已经设计了许多测定方法来测量凋亡过程中最早和最新的步骤。当结合多色检测同时确定几种凋亡特征时,这些检测尤其有效。流式细胞术的多参数特性使该技术特别适合于测量细胞凋亡。在本单元中,我们将介绍分析凋亡细胞中caspase活性的四种主要技术,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分析是评估细胞死亡的有力技术。©2016 by John Wiley &儿子,Inc。
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Measurement and Characterization of Apoptosis by Flow Cytometry

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.

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来源期刊
Current Protocols in Cytometry
Current Protocols in Cytometry Health Professions-Medical Laboratory Technology
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期刊介绍: Published in affiliation with the International Society for Advancement of Cytometry, Current Protocols in Cytometry is a "best practices" collection that distills and organizes the absolute latest techniques from the top cytometry labs and specialists worldwide. It is the most complete set of peer-reviewed protocols for flow and image cytometry available.
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Issue Information Detection and Sorting of Extracellular Vesicles and Viruses Using nanoFACS Live Imaging of the Lung Small Particle Fluorescence and Light Scatter Calibration Using FCMPASS Software. Issue Information
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