Material science provides a direct route to developing a new generation of non-toxic, surface effect-based antifouling technologies with applications ranging from biomedical science to marine transport. The surface topography of materials directly affects fouling resistance and fouling removal, the two key mechanisms for antifouling technologies. However, the field is hindered by the lack of quantified surface characteristics to guide the development of new antifouling materials. Using a biomimetic approach, key surface parameters are defined and quantified and correlated with fouling resistance and fouling removal from the shells of marine molluscs. Laser scanning confocal microscopy was used to acquire images for quantitative surface characterisation using three-dimensional surface parameters, and field assays correlated these with fouling resistance and fouling release. Principle component analysis produced a major component (explaining 54% of total variation between shell surfaces) that correlated with fouling resistance. The five surface parameters positively correlated to increased fouling resistance were, in order of importance, low fractal dimension, high skewness of both the roughness and waviness profiles, higher values of isotropy and lower values of mean surface roughness. The second component (accounting for 20% of variation between shells) positively correlated to fouling release, for which higher values of mean waviness almost exclusively dictated this relationship. This study provides quantified surface parameters to guide the development of new materials with surface properties that confer fouling resistance and release.