Fig 4. Permeation of the Acanthamoeba Neff to the vital dye SYTOX® green caused by addition of IC90 of the bioactive molecules.
Molecules were added to cells (105 cells/ml) in the presence of 1 μM SYTOX® green in PBS + 20 mM d-glucose and the increase in fluorescence (λexc = 485 nm, λem = 520 nm) monitored. Negative Control cells: cells labeled with the Sytox green in presence of 0.5% Methanol. Positive control contained 2.5% Triton X-100 (A). Confocal microscopy of Acanthamoeba castellanii Neff labeled with SYTOX® Green. Amoeba were plated as above and incubated for 3 h with IC90 of the vanillin (B), vanillic acid (C) syringic acid (D) and ursolic acid (E), Negative control (F). Cells were observed in a Leica TSC SPE- confocal microscope equipped with inverted optics (λexc = 482 nm and λem = 519 nm).