Background and Objectives Platelet concentrates are contaminated with residual leucocytes and may also be infected with viruses and bacteria. We investigated whether these pathogens can be inactivated by a two-step procedure comprising photodynamic treatment in the presence of the phenothiazine dye, thionine, followed by irradiation with ultraviolet light (UV-B, wavelength range 290-330 nm). Materials and Methods Platelet concentrates were prepared from buffy coats. The concentrates were spiked with different viruses, bacteria and leucocytes, then illuminated with yellow light in the presence of thionine at dye concentrations between 1 and 5 mum and with UV-B at doses up to 2.4 J/cm(2). The infectivity of samples and the viability of leucocytes were assayed before and after treatment. The influence of treatment on in vitro platelet function was also examined. Results The inactivation of free viruses in platelet concentrates by photodynamic treatment with thionine/light was significantly enhanced when it was followed by irradiation with UV-B. The inactivation of leucocytes and of bacteria by UV-B was improved when it was preceded by thionine/light. Sterile platelet concentrates were prepared from buffy coats infected with Staphylococcus epidermidis . Platelet function and the storage stability of platelet concentrates were only moderately influenced by the two decontamination steps. Conclusions Photodynamic treatment in the presence of the phenothiazine dye, thionine, followed by low-dose UVB, has the potential to inactivate viruses, leucocytes and bacteria, which might contaminate platelet concentrates. Both treatments complement each other.