The first approach to cytometric comparison of cell membrane lipid compositions was that taken by several groups in the 1970's, using diphenylhexatriene (DPH) as a probe and measuring membrane fluidity by polarization as described by Shinitzky and Inbar. In principle, you could play that game with almost any lipophilic probe, e.g., one of the cyanines, getting away from the necessity to use UV excitation, as is required for DPH. Molecular probes sells a number of variants of DPH, most UV-excited, with different membrane binding characteristics. Fluorescence polarization assays are probably not a wonderful way of comparing lipids in different cell types, but would probably serve to discriminate fibroblasts with substantially different membrane cholesterol. Some references: Shinitzky M, Inbar M: Microviscosity parameters and protein mobility in biological membranes. Biochim Biophys Acta 433:133, 1976 Arndt-Jovin D, Ostertag W, Eisen H et al: Studies of cellular differentiation by automated cell separation. Two model systems: Friend virus-transformed cells and Hydra attenuata. J Histochem Cytochem 24:332, 1976 Fox MH, Delohery TM: Membrane fluidity measured by fluorescence polarization using an EPICS V cell sorter. Cytometry 8:20, 1987 Masuda M, Kuriki H, Komiyama Y, Nishikado H, Egawa H, Murata K: Measurement of membrane fluidity of polymorphonuclear leukocytes by flow cytometry. J Immunol Methods 96:225, 1987 Grogan W McL, Collins JM: Guide to Flow Cytometry Methods. New York, Marcel Dekker, 1990, x + 228 pp. -Howard
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