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Miccoli L, Szczepaniak C, Dumas D, et al. 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. J Fluoresc. 1993;3(4):251-5doi: 10.1007/BF00865273.
Miccoli, L., Szczepaniak, C., Dumas, D., Savonnière, S., Muller, S., Carré, M. C., & Donner, M. (1993). 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. Journal of fluorescence, 3(4), 251-5. https://doi.org/10.1007/BF00865273
Miccoli, L, et al. "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." Journal of fluorescence vol. 3,4 (1993): 251-5. doi: https://doi.org/10.1007/BF00865273
Miccoli L, Szczepaniak C, Dumas D, Savonnière S, Muller S, Carré MC, Donner M. 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. J Fluoresc. 1993 Dec;3(4):251-5. doi: 10.1007/BF00865273. PMID: 24234905.
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J Fluoresc. 1993 Dec;3(4):251-5. doi: 10.1007/BF00865273.

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.

Journal of fluorescence

L Miccoli, C Szczepaniak, D Dumas, S Savonnière, S Muller, M C Carré, M Donner

Affiliations

  1. Groupe de Recherches sur les Interactions Moléculaires aux Interfaces, INSERM-CO 10, Brabois, 54511, Vandoeuvre les Nancy, France.

PMID: 24234905 DOI: 10.1007/BF00865273

Abstract

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 (<30 min). Therefore, it can be assumed that DPH-PC can be used as a membrane probe.

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