Microbial Fuel Cell is a bio-electrochemical cell wherein the bacterial metabolism aids in electron generation and hence the production of voltage. In a Microbial Fuel Cell the microorganisms are anaerobically cultured i.e. cultured in absence of terminal electron acceptor i.e. oxygen. This results in generation of electrons in free state making them available for transfer in external circuit. The process, thereby, results in development of potential difference between the two electrodes i.e. anode and cathode. This study involved the extensive study of Microbial Fuel Cell where E.coli was cultured in 3L L.B. media. The voltage recorded was 745mV. The working volume of MFC to 600ml and connected 9 of them in series for voltage upgradation was also studied. The maximum voltage then recorded was 4.5V. In series connection of MFCs also exploited the potential of E.coli cultured in L.B. media. To explore the potentials of microorganisms beyond E.coli to be used in MFC, MFCs based on S.cerevisiae (cultured in Potato Dextrose Broth) and A.niger (cultured in Saboraud’s broth) were developed. These MFCs were to test the potential of other microbial species. Physical parameters like the type of electrode, electrode surface area etc. influences the electrochemical cells. So, these were tested in case of Microbial Fuel Cells by subjecting it to different operational parameters involving higher inoculation, higher cathode area, higher anode area and extra aeration facilitated at cathode. The results obtained were then comparatively analyzed with a MFC being operated under normal conditions i.e. comparison was made with default.