Generation of chemical concentration gradients in mobile droplet arrays via fragmentation of long immiscible diluting plugs.

We report a one-step passive microfluidic technique to generate arrays of moving droplets containing variation of chemical concentration between individual drops. We find that a concentration gradient can be established in a long diluting plug by on-chip dilution and extraction of samples via orthogonal coalescence of the plug with a static array of sample drops. The diluting plug containing the gradient is subsequently fragmented by a droplet generator. We show that the technique is flexible, as the dilution range can be tuned by a variety of control parameters including the carrier fluid flow rate, volume of diluting plugs, and stationary drops. We also find that the concentration gradients have a fine resolution and are reproducible to within 2% relative standard deviation. As one demonstrative application, we show the suitability of the technique for generating a dose-response curve for an enzyme inhibition assay. Because of the ability to inject multiple plugs, our technique has the potential for unlimited as well as sequential dilution of a series of substrates. Thus, our method could be valuable as a high-throughput and high-resolution screening tool for assays that require interrogation of the response of one or more target species to numerous distinct chemical concentrations.