Miniature microbial fuel cells have recently drawn lots of attention as portable power generation devices due to their short startup time and environmentally-friendly process which could be used for powering small integrated biosensors. We designed and fabricated a microbial fuel cell in a microfluidic platform. The device was made in polydimethylsiloxane with a volume of 4 μL and consisted of two carbon cloth electrodes and proton exchange membrane. Shewanella Oneidensis MR-1 was chosen to be the electrogenic bacterial strain and inoculated into the anode chamber. Ferricyanide was used as the catholyte and pumped into the cathode chamber at a constant flow rate during the experiment. The mi- niature microbial fuel cell generated a maximum current of 2.59 μA and had a significantly short startup time.