Qiu Bo, Simon M Celeste
Abramson Family Cancer Research Institute, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Department of Cancer Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Bio Protoc. 2016 Sep 5;6(17). doi: 10.21769/BioProtoc.1912.
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 ., 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 4,4-difluoro-1,3,5,7,8-pentamethyl-4-bora-3a,4a-diaza-s-indacene (BODIPY 493/503) to facilitate quantification of neutral lipid content by flow cytometry and observation of LDs by microscopy.
脂滴（LDs）是普遍存在的动态细胞器，作为中性脂质的储存库，包括甘油三酯和胆固醇酯（Walther和Farese，2012）。脂质种类进出脂滴的运动影响多种细胞过程，如能量稳态、基于脂质的信号传导和膜稳态（Greenberg等，2011）。例如，脂质种类合成或摄取增加时，中性脂质储存会增强。另一方面，细胞外信号可增强包装在中性脂滴内的脂质种类的释放。因此，涉及脂质代谢的课题研究可能需要评估细胞中性脂质含量。在本方案中，我们描述了使用荧光中性脂质染料4,4-二氟-1,3,5,7,8-五甲基-4-硼-3a,4a-二氮杂-s-茚（BODIPY 493/503）通过流式细胞术促进中性脂质含量的定量以及通过显微镜观察脂滴。
