Use of the atomic force microscope to determine the effect of substratum surface topography on the ease of bacterial removal

The ease of removal of differently sized and shaped bacteria from substrata with defined surface topographies and features was investigated. Surfaces with defined surface topography (smooth or with randomly spaced surface features (pits) of 0.5 μm diameter), chemistry (titanium oxide), and wettability (89–93°) were produced. Atomic force microscopy (AFM) was used to determine the ease of bacterial removal from substrata; gram negative Pseudomonas aeruginosa (rods 1 μm width × 3 μm length) and gram positive Staphylococcus aureus (1 μm diameter coccus). The AFM tip was scanned across the retained cells under liquid (contact mode). Over time, using a continuous perpendicular tip force, approximately one third of the cells were removed from the surface following lateral movement of the AFM tip across the surface. When the perpendicular tip force was increased S. aureus were removed more easily from smooth surfaces. In contrast P. aeruginosa cells were removed more easily from the 0.5 μm featured surfaces. The shape of the cell with respect to the shape of the substratum features influences the ease of removal of the cell from the surface: on smooth surfaces the cocci had a smaller cell:surface contact area, whereas the rods had a larger cell:surface contact area. Conversely on featured surfaces the cocci had a larger cell:surface contact area, whereas rods that lay across features had a smaller cell:surface contact area. Using engineered surfaces with defined properties, it has been shown that manipulation of a single parameter (surface roughness) had an effect on the strength of microbial retention.