When a concentrated salt solution and a diluted salt solution are separated by an ion-selective membrane, cations and anions would diffuse at different rates depending on the ion selectivity of the membrane. The difference of positive and negative charges at both ends of the membrane would produce a potential, called the diffusion potential. Thus, electrical energy can be converted from the diffusion potential through reverse electrodialysis. This study demonstrated the fabrication of an energy conversion microchip using the standard micro-electromechanical technique, and utilizing Nafion junction as connecting membrane, which was fabricated by a surface patterned process. Through different salinity gradient of potassium chloride solutions, we experimentally investigated the diffusion potential and power generation from the microchip, and the highest value measured was 135 mV and 339 pW, respectively. Furthermore, when the electrolyte was in pH value of 3.8, 5.6, 10.3, the system exhibited best performance at pH value of 10.3; whereas, pH value of 3.8 yielded the worst.