A new analytical design of continuously-fed microbial fuel cell was built in triplicate in order to investigate relations and effects of various operating parameters such as flow rate and substrate supply rate, in terms of power output and chemical oxygen demand (COD)   removal efficiency. This novel design enables the microbial fuel cell (MFC) systems to be easily adjusted in situ   by changing anode distance to the membrane or anodic volume without the necessity of building many trial-and-error prototypes for each condition. A maximum power output of 20.7 ± 1.9 µW was obtained with an optimal reactor configuration; 2 mM acetate concentration in the feedstock coupled with a flow rate of 77 mL h−  1, an anodic volume of 10 mL and an anode electrode surface area of 70 cm2 (2.9 cm2 projected area), using a 1 cm anode distance tofrom the membrane. COD removal almost showed the reverse pattern with power generation, which suggests trade-off correlation between these two parameters, in this particular example. This novel design may be most conveniently employed for generating empirical data for testing and creating new MFC designs with appropriate practical and theoretical modelling.