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Surface-bound self-assembled lipid nanotubes (LNTs) made of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) were used to visualize the contractile activity of spreading cells. The interaction of cells with LNTs resulted in the nucleation of new nanotubes, directed toward the cell center, from existing ones. This process depended on cell generated forces and required acto-myosin mediated contractility. The dynamics of de novo generation of LNTs upon cell spreading was captured using optical microscopy on fluorescently labeled nanotubes and revealed characteristic fingerprints for different cell types such as fibroblasts, endothelial and melanoma cells. Additionally, the method was applied to detect the effect of a specific inhibitor on the generation of cellular forces. The mechanism of the LNT–cell interaction and the potential applications are discussed. |
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A simultaneous optical waveguide lightmode spectroscopy (OWLS) and electrochemical impedance spectroscopy (EIS) measurement was carried out for the investigation of a supported lipid bilayer and its interactions with a pore-forming peptide, melittin. It was achieved only after the optimization of the ITO coating on the waveguide to increase the electrical sensitivity and the functionalization of the waveguide with a polyelectrolyte to form a lipid bilayer over the ITO surface. The combined system enabled monitoring of melittin pore activities in a wider range of melittin concentrations than either technique alone (1 μg/ml < Cmelittin < 200 μg/ml). Furthermore, it provided unique information that could not be obtained by the individual methods, such as a better identification of the melittin-pore formation and an insight about the correlation between the total pore area vs. adsorbed amount of melittin. |