For the production of ethanol from freshwater cyanobacteria, a new pretreatment method using supercritical fluid was introduced. In this study, it was found that the supercritical fluid could penetrate inside the cell wall and help to liberate starch from cyanobacterial cells which resulted in the increase of the efficiency of ethanol production. For Microcystis aeruginosa, supercritical fluid pretreatment increased the amount of ethanol produced from cyanobacteria from 1.53 g/L to 2.66 g/L. For Anabaena variabilis, the amount of ethanol was increased from 1.25 g/L to 2.28 g/L. With use of supercritical fluid pretreatment, the efficiency of the process to obtain higher ethanol yields from freshwater cyanobacteria was improved upto 80%. The optimum temperature and pressure conditions for supercritical fluid pretreatment were determined as the temperature of $40^{circ}C$ and the pressure of 120 atm. This study demonstrates the feasibility of using supercritical fluid pretreatment for ethanol production using freshwater cyanobacteria.