Most of the implantable medical devices are prone to infection caused by microorganisms that form biofilms. Pseudomonads are frequently used as model species for studying bacterial adhesion. The initial stages of biofilm formation are influenced by different factors including, among others, the chemistry of the surface, the roughness, and topography. The aim of this work was to assess the early stages of Pseudomonas fluorescens biofilm formation on sub-microstructured surfaces (SMS) that are in tune with bacterial size. Copper and gold were used as the substrata. It was concluded that SMS influenced bacterial length, alignment, and distribution, whereas the chemistry of the surfaces affected bacterial length and distribution. However, the effect of the SMS was the most significant. The shape of the bacterial colonies and the polymeric substances production were also influenced by SMS and the chemistry of the surface and both factors may be considered to reduce the susceptibility of a surface to biofilm formation.