细胞死亡的方式主要有两种，包括细胞坏死和细胞凋亡。细胞坏死是早已被认识到的细胞死亡方式，而细胞凋亡是近年来逐渐被认识且越来越受到重视的细胞死亡方式。两种死亡方式最重要的区别是细胞坏死会释放出细胞内溶物，引起炎症反应，而细胞凋亡不会暴露细胞内溶物，一般被吞噬细胞吞噬清除，不引起炎症反应。
细胞凋亡，也称细胞程序性死亡，是指在一定的生理或者病理条件下，细胞主动的、高度有序的、自己结束其生命的过程。细胞凋亡是生物体中一种普遍存在的现象，胚胎形成、个体发育、衰老和损伤细胞的清除等都与细胞凋亡密切相关。细胞凋亡在免疫学中也非常重要，如T细胞在胸腺内发育的过程，经过阴性选择和阳性选择后，95%的胸腺细胞发生凋亡，只有5%的胸腺细胞发育为成熟T细胞进入外周。
检测细胞凋亡的方法很多，如电子显微镜或者光学显微镜下的形态学观察、细胞DNA提取物的DNA梯状带电泳实验等。而流式细胞术是非常重要的检测细胞凋亡的方法，不仅可以定性，也可以定量。流式细胞术检测细胞凋亡的方法也有很多，其中最重要是annexinV/PI双染色法。其他的还有SYTO/PI双染色法、细胞DNA含量分析法、线粒体
损伤检测法、活化的caspase-3检测法、FLICA标记法、甲酰胺诱导ssDNA单抗检测法、TUNEL法等。
标记方法与常规标记荧光抗体的方能一样：加入适量的FITC-annexinV和PI，4℃静置30min即可。注意标记PI的方法与PI染色检测细胞内DNA含量的方法不同，不需提前固定细胞，因为本方法标记PI是为了检测细胞膜的通透性从而鉴别细胞是活细胞还是死细胞，而用PI检测DNA含量时必须先破坏活细胞的细胞膜的完整性使PI进入细胞内与DNA结合。
南京巴傲得生物科技有限公司 ( Biogot technology, co, Ltd），是专一服务于生命科学研究的专业技术型企业。是一大批志同道合的资深专业人士共同创办的具有现代化经营管理水平的生物高科技企业，从事生物技术产品的开发及销售，致力于将巴傲得生物打造成中国乃至世界一流的生物技术产品供应商。巴傲得生物长期备有Bioworld抗体现货8000多种抗体可以在一周左右送到客户的手里。精湛品质、合理价格、专业服务，是我们为您服务的宗旨。在细胞信号通路、免疫学、蛋白组学上拥有显著优势，其抗体产品、生长因子等为生命科学科研工作者提供了极大的便利。客户的满意、微笑和信任是对我们最大的鼓舞与激励，巴傲得生物愿以专业的知识、卓越的服务作为您事业腾飞的基石！
There are two main ways of cell death, including cell necrosis and apoptosis. Cell necrosis has long been recognized as a mode of cell death, while apoptosis has been gradually recognized and paid more and more attention in recent years. The most important difference between the two ways of death is that cell necrosis will release intracellular solutes and cause inflammatory reaction, while apoptosis will not expose intracellular solutes, which are generally phagocytized and eliminated by phagocytes without causing inflammatory reaction.
Apoptosis, also known as programmed cell death, refers to the active, highly orderly and self-ending process of cells under certain physiological or pathological conditions. Apoptosis is a universal phenomenon in living organisms, which is closely related to embryo formation, individual development, aging and elimination of damaged cells. Apoptosis is also very important in immunology, such as the development of T cells in thymus. After negative selection and positive selection, 95% of thymocytes apoptosis, only 5% of thymocytes develop into mature T cells and enter the periphery.
There are many methods to detect apoptosis, such as morphological observation under electron microscope or optical microscope, DNA ladder electrophoresis experiment of cell DNA extract, etc. Flow cytometry is a very important method to detect apoptosis, which can be used not only qualitatively but also quantitatively. There are many methods to detect apoptosis by flow cytometry, the most important of which is annexinV/PI double staining. Others include SYTO/PI double staining, cell DNA content analysis, mitochondrial damage detection,
activated caspase-3 detection, FLICA labeling, formamide-induced ssDNA monoclonal antibody detection, TUNEL method, etc.
The labeling method is the same as the conventional labeling method of fluorescent antibody: add a proper amount of FITC-annexinV and PI, and let stand at 4℃ for 30min. Note that the method of labeling PI is different from the method of detecting DNA content in cells by PI staining, and there is no need to fix cells in advance, because the method of labeling PI is to detect the permeability of cell membrane to identify whether cells are living cells or dead cells, and when using PI to detect DNA content, the integrity of cell membrane of living cells must be destroyed first, so that PI can enter cells to combine with DNA.
Bioworld technology co., Ltd manufactures and supplies antibodies for worldwide distribution. We provide antibodies in bulk quantity and OEM basic services. The experienced scientific and technical professionals make Bioworld Technology a highly innovative, first-class biotechnological company on both manufacturing and marketing. Bioworld strives for a national leader and the world-class company in manufacturing biological products for industry, health, and research. We look forward to serving your research needs and becoming your long-term research partner.
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