Pietkiewicz Sabine, Schmidt Jörn H, Lavrik Inna N
Department of Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University, Magdeburg, Germany.
J Immunol Methods. 2015 Aug;423:99-103. doi: 10.1016/j.jim.2015.04.025. Epub 2015 May 11.
Precisely identifying the type of programmed cell death is one of the key questions in contemporary biomedical research. We developed a straightforward approach allowing quantitative discrimination between two types of cell death on the single cell level: apoptosis and necroptosis. This method uses the combination of imaging flow cytometry with classical Annexin V/propidium iodide staining, which allows for the ascertainment of typical features of dying cells: exposure of the phospholipid phosphatidylserine and the loss of membrane integrity. Image-based analysis of nuclear morphology enables us to distinguish between secondary necrotic/late apoptotic and necroptotic cells directly in one assay. This is a major advantage compared to other contemporary approaches of necroptosis detection, which require a parallel application of several methods. This approach can be used for the quantitative assessment of cell death in cell and systems biology studies of signal transduction networks.
准确识别程序性细胞死亡的类型是当代生物医学研究的关键问题之一。我们开发了一种直接的方法，能够在单细胞水平上对两种细胞死亡类型进行定量区分：细胞凋亡和坏死性凋亡。该方法将成像流式细胞术与经典的膜联蛋白V/碘化丙啶染色相结合，从而能够确定垂死细胞的典型特征：磷脂酰丝氨酸的暴露和膜完整性的丧失。基于图像的核形态分析使我们能够在一次检测中直接区分继发性坏死/晚期凋亡细胞和坏死性凋亡细胞。与其他当代坏死性凋亡检测方法相比，这是一个主要优势，其他方法需要同时应用多种方法。这种方法可用于信号转导网络的细胞和系统生物学研究中细胞死亡的定量评估。
