•Algal hydrophobicity and bubble size are key factors for microalgae flotation.•Algal hydrophobicity can be improved using cationic surfactants at appropriate pHs.•A step-wise optimization of algae flotation is demonstrated. Microalgae harvesting by air flotation is a promising technology for large-scale production of biofuel, feed and nutraceuticals from algae. With an adherence-to-hydrocarbon method and two different types of flotation cells (mechanically agitated cell and Jameson cell), microalgal surface hydrophobicity and bubble size were identified to be critical for effective froth flotation of microalgae. Freshwater alga Chlorella sp. BR2 showed naturally a high hydrophobicity and an ideal response to flotation. However, many marine microalgae possess a low surface hydrophobicity and are thus difficult to harvest. This paper shows that a step-wise optimization approach can substantially improve the flotation of a low surface hydrophobicity marine microalga, Tetraselmis sp. M8, to near full recovery with an enrichment ratio of 11.4.