Curcumin has potential as a photosensitiser (PS) in photodynamic therapy (PDT) for localised superficial infections. However, it is a challenge to make an optimal curcumin formulation in which curcumin has acceptable solubility and stability at physiological pH and combined with high selective phototoxic activity towards bacteria. In the present study, the phototoxic effects of curcumin against gram-negative and gram-positive bacteria were investigated in selected aqueous preparations. The gram-positive Enterococcus faecalis and Streptococcus intermedius and the gram-negative Escherichia coli were used as bacterial models. The bacteria were exposed to 1-25 microM curcumin solubilised in DMSO, cyclodextrines, liposomes and surfactants known to interfere with membranes. After 30 min incubation the bacteria were irradiated with fluorescent tubes emitting blue light (emission max 430 nm). The irradiance was 17 mW/cm2 and the radiant exposure (light dose) was 0.5-30J/cm2. The bacterial survival was calculated as a percentage compared to controls. Various post-irradiation incubation times were tested. Curcumin’s native fluorescence was exploited in examination of curcumin uptake in or adherence to bacteria by fluorescence microscopy. Changes in post-irradiation incubation time, curcumin concentration, irradiation dose and preparation strongly influenced the phototoxic efficiency of curcumin in vitro. Aqueous preparations of DMSO, polyethyleneglycol and the pluronic block copolymer poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) were the most efficient vehicles for curcumin to exert photokilling of gram-positive and gram-negative bacteria.