Culture experiments with phototrophic biofilms taken from the sedimentation tank of the wastewater treatment plant at the Fiumicino Airport in Rome, Italy were carried out in a prototype continuous flow incubator. Biofilms grown at varying photosynthetic photon flux density (PPFD), temperature and flow velocity were sampled at three developmental stages to quantify biofilm dry weight, chlorophyll a concentration and total cellular phosphorus content. While no coherent significant effects by flow and temperature were evidenced, maximum biofilm dry weight and phosphorous concentration significantly increased across all featured PPFDs. Maximum chlorophyll a concentration was saturated above 60 micromol m(-2)s(-1). A highly significant association between organic biomass and phosphorous content was observed for most light conditions, including a larger proportional increase of phosphorus concentration with respect to chlorophyll a at high PPFD. Up to 112 mg P m(-2)d(-1) maximal phosphorous removal rates were achieved. Elemental analysis by energy filtering transmission electron microscopy showed subcellular localization of phosphorus, confirming the accumulation in phototrophic microorganisms in biofilms grown in high light conditions.