In situ imaging of fluid dynamics and nanocarrier nucleation inside microfluidic mixing devices

self-assembly of liposomes under varying flow conditions within a serpentine micromixer. This integrative approach enabled real-time spatial resolution of nanocarrier formation within the microfluidic device, confirming that vesicle formation predominantly occurs at the interface between the aqueous and ethanolic phases, underscoring the robustness of the setup. Beyond advancing mechanistic insight, the complementary use of two confocal microscopy techniques and a Foerster resonance energy transfer-based method offers a powerful toolset for process optimization and in-process quality control. Coupled with advances in additive manufacturing, this approach paves the way for rational micromixer design and the scalable production of microfluidic nanocarrier-based therapeutics, overcoming limitations and accelerating the industrial large-scale production of nanosized therapeutics.