Microplastics (MPs) exposed to the urban coastal seawater could form biofilms, which facilitate the adsorption and transportation of hazardous contaminants. However, influence of biofilms on the metal adsorption of MPs, especially the co-existence of biofilm and metals on MPs, is still less known. In this study, the adsorption of copper (Cu) on biofilm-coated MPs (BMPs) was visually analyzed and quantified. The results of scanning electron microscopy in combination with energy dispersive X-ray showed that biofilm and metals co-occurred on MPs in seawater. The nanoscale secondary ion mass spectrometry images further exhibited that the distribution of Cu, chlorine (Cl) and biofilm on MP surfaces was highly consistent. Moreover, the adsorption of Cu(II) on BMPs was enhanced as quantified by inductively coupled plasma-mass spectrometer. Furthermore, different species on BMPs with and without Cu were identified, and their potential functions of metal or Cl metabolism were predicted based on KEGG pathway database. Overall, for the first time, this study provides visual and quantified evidences for the enhancement of Cu(II) adsorption on BMPs based on co-localization, and it may shed a light on the development of methodologies for investigating the interaction among MPs, biofilms and pollutants in marine environment.