Microfluidic device for real-time formulation of reagents and their subsequent encapsulation into double emulsions

Emulsion drops are often employed as picoliter-sized containers to perform screening assays. These assays usually entail the formation of drops encompassing discrete objects such as cells or microparticles and reagents to study interactions between the different encapsulants. Drops are also used to screen influences of reagent concentrations on the final product. However, these latter assays are less frequently performed because it is difficult to change the reagent concentration over a wide range with high precision within a single experiment. In this paper, we present a microfluidic double emulsion drop maker containing pneumatic valves that enable injection of different reagents using pulsed width modulation and subsequent mixing. This device can produce drops from reagent volumes as low as 10 μl with minimal sample loss, thereby enabling experiments that would be prohibitively expensive using droplet generators that do not contain valves. We employ this device to monitor the kinetics of cell free synthesis of green fluorescent proteins inside double emulsions. To demonstrate the potential of this device, we perform DNA titration experiments in double emulsion drops to test the influence of the DNA concentration on the amount of green fluorescence proteins produced.