Resolution of plasma membrane lipid fluidity in intact cells labelled with diphenylhexatriene.
The partitioning of fluorescence probes into intracellular organelles poses a major problem when fluorescence methods are applied to evaluate the fluidity properties of cell plasma membranes with intact cells. This work describes a method for resolution of fluidity parameters of the plasma membrane in intact cells labelled with the fluorescence polarization probe 1,6-diphenyl-1,3,5-hexatriene (DPH). The method is based on selective quenching, by nonradiative energy transfer, of the fluorescence emitted from the plasma membrane after tagging the cell with a suitable membrane impermeable electron acceptor. Such selective quenching is obtained by chemical binding of 2,4,6-trinitrobenzene sulfonate (TNBS), or by incorporation of N-bixinoyl glucosamine (BGA) to DPH-labelled cells. The procedures for determination of lipid fluidity in plasma membranes of intact cells by this method are simple and straightforward.
D000818:Animals; D002462:Cell Membrane / Q000502:physiology*; D002478:Cells, Cultured; D004161:Diphenylhexatriene; D004735:Energy Transfer; D004910:Erythrocyte Membrane / Q000502:physiology; D006801:Humans; D008560:Membrane Fluidity; D008563:Membrane Lipids / Q000502:physiology*; D051379:Mice; D011090:Polyenes; D013050:Spectrometry, Fluorescence; D013574:Synaptosomes / Q000502:physiology
D008563:Membrane Lipids; D011090:Polyenes; D004161:Diphenylhexatriene