Time-of-flight secondary ion mass spectrometry (ToF-SIMS) has been employed to study the biofouling of stainless steel samples immersed in seawater. The aim of these characterisations was to understand the initial mechanisms of biomolecule adsorption for relatively short immersion times (from 0 to 24 h).The results show that: (i) there were unavoidable sample ''precontaminations'' on the surfaces, despite precaution during their preparation and manipulation (washing, drying and storing); (ii) the major peaks detected were the substrate ones whatever the immersion time [However, some organic (nitrogen and oxygen containing) and inorganic secondary ions appeared and grew with the immersion time.]; (iii) the surface contaminations, the nonuniformity of the adsorbed material so as and bacteria have been clearly observed by high-lateral resolution molecular ToF-SIMS mapping.