4.5. Atomic Force Microscopy (AFM)

Cells for AFM were established in 60 mm Petri plates in the presence and absence of sub-inhibitory concentrations of different EOs. After 48 h of incubation, the liquid medium was withdrawn, and the plates were washed twice with PBS, the samples fixed with 2.5% glutaraldehyde at 4 °C for 4 h, washed with distilled water and air dried [64]. All images were collected with a Wet-SPM-9600 scanning probe microscope (Shimadzu, Kyoto, Japan) in contact mode using Si₃N₄ cantilevers. The scanning area in the images was 5 × 5 μm with at least 20 different cells imaged for each biological replicate.