title: Metabolic Assays for Detection of Neutral Fat Stores.
abstract: Lipid droplets (LDs) are ubiquitous intracellular structures whose formation, growth, and maintenance are highly regulated (Wanget al., 2013; Ranallet al., 2011; Goodman, 2009). Lipid metabolism and droplet dynamics are of considerable interest to agriculture, biofuel production, viral pathology, nutrition, and cancer biology (Walther and Farese, 2009; Liuet al., 2010). Accumulation of fatty acids and neutral lipids in nonadipose tissues is cytotoxic (Kourtidiset al., 2009). BODIPY 493/503 (4,4-Difluoro-1,3,5,7,8-Pentamethyl-4-Bora-3a,4a-Diaza-s-Indacene) is the standard dye to study LDs within adipocytes. BODIPY 493/503 contains a nonpolar structure that, upon binding to neutral lipid, emits a green fluorescence signal with a narrow wavelength range, making it an ideal fluorophore for multi-labeling experiments. The hydrophobic nature of the dye molecules promotes rapid entry into the nonpolar environment of LDs (Listenberge and Brown, 2007). Gocze and Freeman showed that the lipid fluorescent variability is significantly lower when using BODIPY493/503 compared to Nile Red, suggesting that it may be more specific for the LD (Gocze and Freeman, 1994). Here, we describe a BODIPY 493/503 assay for the detection of neural fat stores in cultured cells (Figure 1) (Wanget al., 2013).
authors.affiliation: Cancer Research Center, Department of Biomedical Sciences, University at Albany, State University of New York, New York, USA.
authors.forename: Jan M
authors.initials: JM
authors.lastname: Baumann
authors.affiliation: Cancer Research Center, Department of Biomedical Sciences, University at Albany, State University of New York, New York, USA.
authors.forename: Leila
authors.initials: L
authors.lastname: Kokabee
authors.affiliation: Cancer Research Center, Department of Biomedical Sciences, University at Albany, State University of New York, New York, USA.
authors.forename: Xianhiu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Cancer Research Center, Department of Biomedical Sciences, University at Albany, State University of New York, New York, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Cancer Research Center, Department of Biomedical Sciences, University at Albany, State University of New York, New York, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Wong
authors.affiliation: Cancer Research Center, Department of Biomedical Sciences, University at Albany, State University of New York, New York, USA.
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Conklin