Microalgae lipid extracted by organic solvent is a solid–liquid extraction process. Therefore, lipid extraction yield and extraction rate are notably affected by the diffusion rate of lipids between the solid–liquid phase. In the present study, lipid yields of single and tertiary cross flow extraction by EA/E (1:1, v/v) were compared. It was found that tertiary extraction obtained lipid yield of 6.16%, compared to that of single extraction of only 4.62%, with a total lipid yield improvement of 33.3% with the same solvent consumption, which could be attributed to greater lipid concentration gradient between solid and liquid phase in the second and third stage of tertiary cross flow extraction, which enhanced the lipids transfer from microalgal cell to the solvent. Lower lipid yield of first stage of tertiary cross flow extraction process than that of single extraction process was observed since only one third of solvent was used as indicated in Section 3.2. However, the tertiary process is more complicated than single extraction process. Further study should be conducted to analyze relationship between lipid yield and economic feasibility for lipid extraction by EA/E.