Aqueous Two-Phase System Formation in Small Droplets by Shirasu Porous Glass Membrane Emulsification Followed by Water Extraction

By utilizing water transport phenomena between two different water-in-oil (W/O) emulsion droplets through continuous oil phase, we developed a novel method of aqueous two-phase system (ATPS) formation in small droplets prepared by Shirasu porous glass (SPG) membrane emulsification technique. When we mixed W/O emulsion droplets containing poly(ethylene glycol) (PEG) and dextran (DEX) at concentrations below the threshold of the phase separation, with droplets containing other solutes at high concentrations, water extraction from the droplets containing PEG and DEX to those containing the other solutes occurred, owing to the osmotic pressure difference. This effect increased the concentrations of PEG and DEX in the droplets above the phase separation threshold. We demonstrated the feasibility of the preparation method by varying the pore sizes of the SPG membranes, the solutes, and their concentrations. Only when the concentration of the solute was high enough to extract sufficient amounts of water did the homogeneous disperse phase consisting of PEG and DEX in droplets turn into a PEG-rich phase and DEX-rich phase, showing ATPS. This result was irrespective of the solute itself and pore size of the SPG membrane. In particular, we successfully demonstrated monodisperse ATPS droplets with diameters of approximately 10 mu m under a certain condition.