The European Interreg Italy–France 2014–2020 Maritime Project SPlasH! (Stop to Plastics in H₂O!) focused on the study of microplastics (MPs) in the marine port environment to evaluate their presence, abundance, and mechanisms of diffusion to the open sea. In the framework of this project, a worldwide review of 74 studies was carried out, providing an overview of MP investigation techniques, focusing on sampling strategies, laboratory methodologies, and identification of MPs collected in seawater, and specifically evaluating their applicability to the marine port environment. Nets were the most commonly used device for MP surface sampling, but their use can be difficult in narrow spaces within the port basins, and they must be coupled to discrete sampling devices to cover all port basins. In the laboratory, density separation (NaCl, ZnCl₂, NaI, sodium lauryl sulfate (SLS)), filtration (polycarbonate, polyamide, glass, cellulose, ANOPORE inorganic membrane filters), sieving, visual sorting, and digestion methods (acidic, enzymatic, alkaline, oxidative) were used to separate MPs from seawater. Digestion becomes essential with water samples with great inorganic and organic loads as deriving from a port. Although many studies are based only on visual MP identification under a microscope, analytical identification techniques unequivocally determine the particle nature and the identity of the plastic polymers and are necessary to validate the visual sorting of MPs. Fourier-transform infrared spectroscopy (FTIR) is the most used analytical identification technique.