Recent progress in the synthesis of all-aqueous two-phase droplets using microfluidic approaches

An aqueous two-phase system (ATPS) is a system with liquid-liquid phase separation and shows great potential for the extraction, separation, purification, and enrichment of proteins, membranes, viruses, enzymes, nucleic acids, and other biomolecules because of its simplicity, biocompatibility, and wide applicability [1-4]. The clear aqueous-aqueous interface of ATPSs is highly advantageous for their implementation, therefore making ATPSs a green alternative approach to replace conventional emulsion systems, such as water-in-oil droplets. All aqueous emulsions (water-in-water, w-in-w) hold great promise in the biomedical field as glucose sensors [5] and promising carriers for the encapsulation and release of various biomolecules and nonbiomolecules [6-10]. However, the ultralow interfacial tension between the two phases is a hurdle in generating w-in-w emulsion droplets. In the past, bulk emulsification and electrospray techniques were employed for the generation of w-in-w emulsion droplets and the fabrication of microparticles and microcapsules in the later stage. Bulk emulsification is a simple and low-cost technique; however, it generates polydisperse w-in-w emulsion droplets. Another technique, electrospray, involves easy experimental setups that can generate monodisperse but nonspherical w-in-w emulsion droplets. In comparison, microfluidic platforms provide monodisperse w-in-w emulsion droplets with spherical shapes, deal with the small volumes of solutions and short reaction times and achieve portability and versatility in their design through rapid prototyping. Owing to several advantages, microfluidic approaches have recently been introduced. To date, several different strategies have been explored to generate w-in-w emulsions and multiple w-in-w emulsions and to fabricate microparticles and microcapsules using conventional microfluidic devices. Although a few review articles on ATPSs emulsions have been published in the past, to date, few reviews have exclusively focused on the evolution of microfluidic-based ATPS droplets. The present review begins with a brief discussion of the history of ATPSs and their fundamentals, which is followed by an account chronicling the integration of microfluidic devices with ATPSs to generate w-in-w emulsion droplets. Furthermore, the stabilization strategies of w-in-w emulsion droplets and microfluidic fabrication of microparticles and microcapsules for modern applications, such as biomolecule encapsulation and spheroid construction, are discussed in detail in this review. We believe that the present review will provide useful information to not only new entrants in the microfluidic community wanting to appreciate the findings of the field but also existing researchers wanting to keep themselves updated on progress in the field.