Microfluidic Platform for Intracellular Liquid-liquid Phase Separation Studies

Membraneless organelles (MLOs) formed through liquid-liquid phase separation (LLPS) are becoming increasingly relevant to understand viral-host interactions, neurodegenerative disease, and cancer. The modulation of LLPS involves many parameters and components. To describethesemodulators, typical in vitro studiesrequirelaborious, manual samplepreparation of different concentrations and costly biological reagents. In this thesis, a minimal reagent, microfluidic platform is developed to systematically generate samples of different concentrations and trigger phase separation. The platform’s utility is tested by constructing phase diagrams describing the modulation of LLPS using an aqueous two-phase system (ATPS) and an MLO-based phase separating system. Then, on-chip biophysical characterization typical of in vitro studies is conducted. This platform will be utilized by scientists to study the growing number of MLOs and inform clinical treatments for pathology related to LLPS.