Interaction of a phosphatidylcholine derivative of 1,6-diphenyl-1,3,5-hexatriene (DPH) with intact living cells: Steady-state fluorescence polarization and phase fluorometry studies
Autor: | Sylvaine Muller, Dominique Dumas, C. Szczepaniak, L. Miccoli, Marie-Christiane Carré, Mireille Donner, S. Savonnière |
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Rok vydání: | 1993 |
Předmět: |
Sociology and Political Science
medicine.diagnostic_test biology Chemistry Clinical Biochemistry Phospholipid Analytical chemistry Biochemistry Fluorescence Fluorescence spectroscopy Flow cytometry Clinical Psychology chemistry.chemical_compound Phospholipase A2 Membrane Phosphatidylcholine medicine biology.protein Biophysics lipids (amino acids peptides and proteins) Anisotropy Law Spectroscopy Social Sciences (miscellaneous) |
Zdroj: | Journal of Fluorescence. 3:251-255 |
ISSN: | 1573-4994 1053-0509 |
DOI: | 10.1007/bf00865273 |
Popis: | The potential interest of DPH-PC was checked with a macrophagic cell line (P388D1). The uptake of DPH-PC was associated with a rapid increase in both fluorescence intensity and a slow decrease in anisotropy values. A flow cytometry comparative study with DPH revealed in both cases the existence of two cell subpopulations with different labeling levels. The analysis of fluorescence decay of DPH-PC showed two components. The fractional intensity of the main component (9.7 ns) is higher than 92%. The Lorentzian distribution of the main lifetime presents an important homogeneity. The observation that an increase in temperature induced a decrease in steady state anisotropy values but did not affect the lifetime suggests that the anisotropy variations effectively reflect modifications in the cohesion of probe micro-surroundings. A transmembrane diffusional phenomenon of a fraction of fluorescent phospholipids (205) was suggested by a study with a nonpermeant membrane quencher. The transmembrane diffusion was confirmed by extraction of the phospholipid analog with fatty acid free BSA. The use of inhibitors of endogenous phospholipase A2 showed a progressive hydrolysis of the fluorescent phospholipid. Nevertheless, the hydrolysis can be neglected in the case of short term interactions with cells ( |
Databáze: | OpenAIRE |
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