Re: membrane lipid probe
Howard Shapiro (hms@shapirolab.com)
Sun, 31 Aug 1997 20:04:22 -0400 (EDT)
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