The temporal resolution of the SAXS system was insufficient to observe structural changes in liposomes during formation, and thus, further development of microfluidic systems combined with synchrotron facilities is required. However, our findings clarified the effects of time-dependent changes in ethanol concentration on liposome structures. Generally, the liposome suspension contains approximately 30% residual ethanol, which is removed during dialysis. Our results suggest that the post-treatment process to remove residual ethanol, in addition to the liposome formation process, is critical. Microfluidic devices focusing on the post-treatment process or integrating liposome production and post-treatment processes may accelerate the development of liposome-based nanomedicine.