To prevent the occurrence of the organic crossover in membraneless microbial fuel cells (ML-MFCs), dual-node MFC (DA-MFC) was designed from multi-anode concept to ensure anode zone. The anode zone addressed increase the utilization of organic matter in ML-MFCs, as the result, the organic crossover was prevented and performance of MFCs were enhanced. The maximum power of the DA-MFC was 0.46 mW, which is about 1.56 times higher than the ML-MFC (0.29 mW). Furthermore, the DA-MFC had advantage in correlation of organic substance concentration and dissolved oxygen concentration, and even electric over-potential. In addition, in terms of cathode fouling, the DA-MFC showed clearer surface. Hence, the anode zone should be considered in the advanced ML-MFC for practically use in wastewater treatment process, and also for scale-up of MFCs. (C) 2016 Elsevier Ltd. All rights reserved.