This review summarizes recent advances of aqueous two-phase systems (ATPSs), particularly their interfaces, with a focus on biomedical applications.

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Digital loop‐mediated isothermal amplification (dLAMP) refers to compartmentalizing nucleic acids and LAMP reagents into a large number of individual partitions, such as microchambers and droplets. This compartmentalization enables dLAMP to be an excellent platform to quantify the absolute number of the target nucleic acids. Owing to its low requirement for instrumentation complexity, high specificity, and strong tolerance to inhibitors in the nucleic acid samples, dLAMP has been recognized as a simple and accurate technique to quantify pathogenic nucleic acid. Herein, the general process of dLAMP techniques is summarized, the current dLAMP techniques are categorized, and a comprehensive discussion on different types of dLAMP techniques is presented. Also, the challenges of the current dLAMP are illustrated together with the possible strategies to address these challenges. In the end, the future directions of the dLAMP developments, including multitarget detection, multisample detection, and processing nucleic acid extraction are outlined. With recently significant advances in dLAMP, this technology has the potential to see more widespread use beyond the laboratory in the future.

Droplets containing ternary mixtures can spontaneously phase-separate into high-order structures upon a change in composition, which provides an alternative strategy to form multiphase droplets. However, existing strategies always involve nonaqueous solvents that limit the potential applications of the resulting multiple droplets, such as encapsulation of biomolecules. Here, a robust approach to achieve high-order emulsion drops with an all-aqueous nature from two aqueous phases by osmosis-induced phase separation on a microfluidic platform is presented. This technique is enabled by the existence of an interface of the two aqueous phases and phase separation caused by an osmolality difference between the two phases. The complexity of emulsion drops induced by phase separation could be controlled by varying the initial concentration of solutes and is systematically illustrated in a state diagram. In particular, this technique is utilized to successfully achieve high-order all-aqueous droplets in a different aqueous two-phase system. The proposed method is simple since it only requires two initial aqueous solutions for generating multilayered, organic-solvent-free all-aqueous emulsion drops, and thus these multiphase emulsion drops can be further tailored to serve as highly biocompatible material templates.

In this study, we develop a method to detect multiple DNAs of foodborne pathogens by encapsulating emulsion droplets for loop-mediated isothermal amplification (LAMP). In contrast to the traditional bulk-phase LAMP, which involves a labor-intensive mixing process, with our method, different primers are automatically mixed with DNA samples and LAMP buffers after picoinjection. By directly observing and analyzing the fluorescence intensity of the resultant droplets, one can detect DNA from different pathogens, with a detection limit 500 times lower than that obtained by bulk-phase LAMP. We further demonstrate the ability to quantify bacteria concentration by detecting bacterial DNA in practical samples, showing great potential in monitoring water resources and their contamination by pathogenic bacteria.

Studying the stability of Pickering emulsion is of great interest for applications including catalysis, oil recovery, and cosmetics. Conventional methods emphasize the overall behavior of bulk emulsions and neglect the influence of particle adsorbing dynamics, leading to discrepancies in predicting the shelf-life of Pickering emulsion-based products. By employing a microfluidic method, the particle adsorption is controlled and the stability of the Pickering emulsions is consequently examined. This approach enables us to elucidate the relationship between the particle adsorption dynamics and the stability of Pickering emulsions on droplet-level quantitatively. Using oil/water emulsions stabilized by polystyrene nanoparticles as an example, the diffusion-limited particle adsorption is demonstrated and investigated the stability criteria with respect to particle size, particle concentration, surface chemistry, and ionic strength. This approach offers important insights for application involving Pickering emulsions and provides guidelines to formulate and quantify the Pickering emulsion-based products.

Purpose: Silicone oil (SiO) with additives of high-molecular-weight (HMW) SiO molecules, eases both the injection and removal. When used inside an eye, it remains unclear how increasing extensional viscosity of SiO might reduce emulsification. Using cell-lined models, this study aims to understand the reason why SiO with HMW is less prone to emulsification. Methods: The adhesion of SiO was studied and recorded in a cell-coated microchannel by optical microscopy. The resistance of SiO against emulsification was tested on another cell-coated eye-on-a-chip platform, which was subject to simulated saccadic eye movements, for 4 days. Silicone oil (SiO) candidates with HMW, SiO HMW2000 and SiO HMW5000 , and their counterparts SiO 2000 and SiO 5000 without HMW, were tested. The quantity of the SiO emulsified droplets formed was assessed daily by optical microscopy. Results: When flowing in the microchannel, SiO adheres on the cell-coated substrate. The number of droplets is generally lower in SiO with HMW than their counterparts. At the end of the experiment, the average numbers of droplets in SiO 2000 (29.1 AE 41.0) and SiO 5000 (9.1 AE 19.5) are higher than those in SiO HMW2000 (6.0 AE 4.5) and SiO HMW5000 (5.6 AE 4.1). Conclusion: A new mechanism of emulsification of SiO is proposed: SiO adheres to ocular tissue to form emulsified droplets. The presence of HMW, which increases the extensional viscosity, may resist the break-up of SiO from the substrate to form emulsified droplets. When tested in a physiologically representative platform, the use of HMW in SiO generally reduces the number of droplets formed in vitro.

We use a glass-based microfluidic device to generate non-equilibrium water-in-water-in-oil (w/w/o) double emulsions and study how they transform into equilibrium configurations. The method relies on using three immiscible liquids, with two of them from the phase-separated aqueous two-phase systems. We find that the transformation is accompanied by an expansion rim, while the characteristic transformation speed of the rim mainly depends on the interfacial tension between the innermost and middle phases, as well as the viscosity of the innermost phase when the middle phase is nonviscous. Remarkably, the viscosity of the outermost phase has little effect on the transformation speed. Our results account for the dynamics of non-equilibrium double emulsions towards their equilibrium structure and suggest a possibility to utilize the non-equilibrium drops to synthesize functional particles.