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Author:
Date:
2016-10-05
[Abstract] The production of free radicals is the result of normal cellular metabolism. Free radicals are involved in innumerable different cellular and biological functions such as signaling, proliferation, cell death, aging, inflammation, etc. Under physiological conditions, the levels of reactive oxygen species (ROS) are strictly regulated by the cells. However, during stressful conditions such as oxidative stress, ROS levels increase causing damages to different molecules like DNA, lipids and proteins. Increased levels of ROS have been associated with a growing list of different diseases. In this protocol, we used MitoSOX and CellROX Green oxidative stress probes to label the intracellular ROS and detect the fluorescence using cell sorting and confocal analyses.
[摘要] 自由基的产生是正常细胞代谢的结果。 自由基涉及无数不同的细胞和生物学功能，例如信号传导，增殖，细胞死亡，衰老，炎症等。 在生理条件下，活性氧（ROS）的水平被细胞严格调节。 然而，在应激条件例如氧化应激期间，ROS水平增加导致对不同分子如DNA，脂质和蛋白质的损伤。 增加的ROS水平与不断增加的不同疾病列表相关联。 在这个协议中，我们使用MitoSOX和CellROX绿色氧化应激探针标记细胞内ROS和使用细胞分选和共聚焦分析检测荧光。
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Author:
Date:
2016-09-05
[Abstract] Lipid droplets (LDs) are ubiquitous, dynamic organelles and function as a storage depot for neutral lipids, including triglycerides and cholesterol esters (Walther and Farese, 2012). The movement of lipid species into and out of LDs impacts a variety of cellular processes, such as energy homeostasis, lipid-based signaling, and membrane homeostasis (Greenberg et al., 2011). For example, neutral lipid storage is enhanced upon increased synthesis or uptake of lipid species. On the other hand, extracellular signals can enhance the release of lipid species packaged within neutral LDs. Thus, the investigation of topics involving lipid metabolism may require the assessment of cellular neutral lipid content. In this protocol, we describe the use of the fluorescent neutral lipid dye ...
[摘要] 脂滴（LD）是普遍存在的，动态的细胞器，并且作为中性脂质包括甘油三酯和胆固醇酯的储存库（Walther和Farese，2012）。 脂质物质进出LD的运动影响多种细胞过程，例如能量稳态，基于脂质的信号传导和膜内环境稳定（Greenberg等人，2011）。 例如，当增加脂质种类的合成或摄取时，中性脂质储存增强。 另一方面，细胞外信号可以增强包装在中性LD内的脂质物质的释放。 因此，涉及脂质代谢的主题的研究可能需要评估细胞中性脂质含量。 在该方案中，我们描述了荧光中性脂质染料4,4-二氟-1,3,5,7,8-五甲基-4-硼-3a，4a-二氮杂-s-引达省（BODIPY 493/503 ），以通过流式细胞术和通过显微镜观察LD来促进中性脂质含量的定量。
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