Bio-removal of heavy metals from wastewater by microalgae has been investigated for decades. However, technical and economical limitations of cultivation systems for microalgae still impair progress toward application. Recently, a novel type of bioreactor for (immobilized) biofilm cultivation, the Porous Substrate Bioreactor (PSBR), has been shown to optimize biomass feedstock production and harvest, offering novel possibilities for application in the treatment of wastewater. We used two types of laboratory-scale Twin-Layer PSBRs to remove zinc (2-3 mg Zn L-1) from synthetic wastewater and real mine dump leachate in a continuous and batch process. The selection and use of a biofilm of a Zn-resistant strain of the green alga Stichococcus bacillaris (EC50 of 28.9 mg Zn L-1 based on Pulse-amplitude modulated (PAM) chlorophyll fluorescence analysis) led to a high zinc absorption capacity of 15-19 mgZn g(-1) algal dry matter. The removal capacity for zinc correlated positively with biomass production and was thus, light dependent. Bio-removal properties observed here combined with biomass productivities of PSBR systems compare favorably with other algal-based bio-sorption technologies. (C) 2015 Elsevier B.V. All rights reserved.