Some species of microalgae have high productivity and lipid content, which makes them good candidates for biodiesel production. Biomass separation and cell disruption are important steps in biodiesel production from microalgae. In this work, we explored the fundamentals of electroflotation by alternating current (EFAC) with non-consumable electrodes to simultaneously harvest microalgae and disrupt cells from mixed microalgae obtained from waste stabilization ponds. The harvesting efficiency was evaluated using chlorophyll-a and turbidity, which reached removals of 99% and 95%, respectively, during a batch time of 140 min. Cell disruption was evaluated using lipid extraction, and the best results were achieved with a batch time of 140 min, which resulted in a 14% yield. Therefore, EFAC was shown to be an attractive potential technology for simultaneous microalgal harvesting and cell disruption.