Microalgae have great potential for biofuel production and wastewater treatment, but the high cost of harvesting hinders their practical application. In this study, economical harvesting of hydrophobic microalgae by electro-flotation without coagulation was assessed. The harvesting performance of this method for selected species of freshwater microalgae with different degrees of hydrophobicity (Tribonema sp., highly hydrophobic; Scenedesmus sp., moderately hydrophobic; and Pandorina sp., hydrophilic) were compared. It was found that microalgal hydrophobicity played a critical role in electro-flotation. Under the same condition (current 0.3 A, velocity gradient 200 s⁻¹, biomass concentration 1 g/L), Tribonema sp. could be effectively harvested (96.2 ± 0.4%) after 20 min of electro-flotation, while the harvesting efficiency decreased significantly with Scenedesmus sp. (70.1 ± 5.2%, 20 min) and Pandorina sp. (<10%, 1 h). The influences of current, electrolysis time, mixing intensity (velocity gradient) and biomass concentration on Tribonema sp. (hydrophobic) harvesting were further investigated. Increasing the current within a certain range (0.1 A-0.4 A) was beneficial to harvesting, while it's further increase decreased floating velocity, which was similar to the effect of the velocity gradient. Under the optimal condition, the harvesting efficiency of Tribonema sp. was 96.3% and the energy consumption (0.19 kWh/kg biomass) was much lower than other harvesting techniques, indicating that electro-flotation is a time-saving and economical approach for hydrophobic microalgae harvesting.