Identification of ROS Using Oxidized DCFDA and Flow-Cytometry
Identification of ROS Using Oxidized DCFDA and Flow-Cytometry
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DOI：
10.1007/978-1-60761-411-1_4
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发表时间：
2010-01-01
期刊：
影响因子：
--
通讯作者：
Kusmartsev, Sergei
中科院分区：
文献类型：
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作者：
Eruslanov, Evgeniy;Kusmartsev, Sergei
细胞在有氧代谢过程中不断产生活性氧（ROS）。ROS的产生在免疫系统中起着重要的保护和功能作用。细胞拥有强大的抗氧化防御系统，以对抗ROS的过度产生。当ROS的产生破坏细胞的天然抗氧化防御时，细胞中发生氧化应激。ROS和氧化损伤被认为在许多人类疾病中起重要作用，包括癌症、动脉粥样硬化、其他神经退行性疾病和糖尿病。因此，确定它们的确切作用需要能够准确测量ROS及其引起的氧化损伤。有许多方法可以测量细胞中自由基的产生。最直接的技术使用细胞可渗透的荧光和荧光探针。2 '-7'-二氯二氢荧光素二乙酸酯（DCFH-DA）是用于直接测量电池的氧化还原状态的最广泛使用的技术之一。它有几个优点比其他技术开发。它非常容易使用，对细胞氧化还原状态的变化非常敏感，价格低廉，可用于跟踪ROS随时间的变化。
Cells constantly generate reactive oxygen species (ROS) during aerobic metabolism. The ROS generation plays an important protective and functional role in the immune system. The cell is armed with a powerful antioxidant defense system to combat excessive production of ROS. Oxidative stress occurs in cells when the generation of ROS overwhelms the cells' natural antioxidant defenses. ROS and the oxidative damage are thought to play ail important role in many human diseases including cancer, atherosclerosis, other neurodegenerative diseases and diabetes. Thus, establishing their precise role requires the ability to measure ROS accurately and the oxidative damage that they cause. There arc many methods for measuring free radical production in cells. The most straightforward techniques use cell permeable fluorescent and chemiluminescent probes. 2'-7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) is one of the most widely used techniques for directly measuring the redox state of a cell. It has several advantages over other techniques developed. It is very easy to use, extremely sensitive to changes in the redox state of a cell, inexpensive and can be used to follow changes in ROS over time.