A programmable microenvironment for cellular studies via microfluidics-generated double emulsions

High throughput cellular studies require small sample volume to reduce costs and enhance sensitivity. Microfluidics-generated water-in-oil (W/O) single emulsion droplet systems, in particular, provide uniform, well defined and discrete microenvironment for cell culture, screening, and sorting. However, these single emulsion droplets are incapable of continuous supply of nutrient molecule and are not compatible with aqueous phase-based analysis. A solution is to entrap W/O droplets in another aqueous phase, forming water-in-oil-in-water (W/O/W) double emulsions. The external aqueous phase efficiently prevents desiccation and reduces the amount of organic component, and yet retaining the advantages of compartmentalization. The internal environment can also be programmed dynamically without the need of rupturing the droplets. In this study, we explore the potential application of W/O/W double emulsion droplets for cell cultivation, genetic activation and study of more complicated biological events such as bacteria quorum-sensing as an example. This study demonstrates the advantages and potential application of double emulsion for the study of complex biological processes.