The new nanotechnologies in biomaterials for cardiovascular applications target at surface alterations for prevention of platelets aggregation and subsequent clotting as their usual failure arises from thrombogenicity. Knowledge of structural properties of platelets during their adhesion on nanostructure materials is required to obtain a comprehensive understanding of their activation and the conventional imaging tools require special preparation of the samples and does not guarantee the viability of the cells. Thus, in this study, the atomic force microscope (AFM) which is a non-destructive and nanoscale precision technique is implemented for the study of platelets' adhesion onto amorphous hydrogenated carbon (a-C:H) thin films and a methodology is developed. Carbon-based thin films grown by magnetron sputtering under different deposition conditions are considered to meet the requirements for biomedical applications and were selected as well-characterized, case study materials. Platelet rich plasma drawn from healthy donors was used for the study of platelets adhesion onto the a-C:H films. The fourier transform IR phase modulated spectroscopic ellipsometry (FTIRSE) (900-3500 cm -1 ) being a powerful, non-destructive, optical technique was used for the investigation of bonding structure of the adherent platelets onto the a-C:H materials and the contribution of the different vibration bands of the platelet bonding groups was shown and discussed. The effect of nanostructure, surface properties and wettability of the carbon thin films on their thrombogenic potential was verified and it was found that the different deposition conditions determine their structural, surface and biological properties. Thus, the tailoring of surface properties of biomaterials and the informative study of platelets-nanomaterials interactions with AFM and FTIRSE will revolutionize the development of less thrombogenic biomaterials.