Microbial biofilms were formed on coupons with defined coatings in once‐through laminar flow fields of controlled bulk‐phase composition and shear. Dilute media were utilized to select for biofilm growth. The formation, succession, and stability of the biofilms were monitored with non‐destructive on‐line methods (fluorescence, bioluminescence, attenuated total reflectance Fourier transform infrared spectrometry [ATR‐FTIR] and electrochemical impedance spectroscopy) and by high resolution destructive analysts (viable and direct counts and phospholipid fatty acid signature methods) at the termination of the experiments. Biofilms of reproducible composition can be formed and the order of inoculation of multi‐component biofilms affects their composition at harvest. The corrosion rates of mild steel depended on the biofilm composition but not the attached biomass. Examination of biofilms with the scanning vibrating electrode in a microscope field showed effects of heterogeneity in biofilm structure which promoted localized anodic activity.Pseudomonasstains were engineered to contain theluxgene cassette as a “reporter”; and the formation of the exopolymer alginate was shown not to promote attachment of the strain or secondary colonization byVibrio. Examination of mutants forming different alginate structures showed differential attachment and biofilm structure. Studies of mutants of lipopolysaccharide structure showed differential attachment to substrata. Specific antifouling and fouling‐release coatings showed a wide range of attachment and release properties as well as sublethal toxicity.