Towards an active droplet-based microfluidic platform for programmable fluid handling

Droplet-based microfluidic systems have emerged as powerful alternatives to conventional high throughput screening platforms, due to their operational flexibility, high-throughput nature and ability to efficiently process small fluid volumes. However, the challenges associated with performing bespoke operations on user-defined droplets often limit their utility in screening applications that involve complex workflows. To this end, the marriage of droplet- and valve-based microfluidic technologies offers the prospect of balancing the controllability of droplet manipulations and analytical throughput. In this spirit, we present a microfluidic platform that combines the capabilities of integrated microvalve technology with droplet-based sample compartmentalization to realize a highly adaptable programmable fluid handling functionality. The microfluidic device consists of a programmable formulator linked to an automated droplet generation device and storage array. The formulator leverages multiple inputs coupled to a mixing ring to produce combinatorial solution mixtures, with a peristaltic pump enabling titration of reagents into the ring with picoliter resolution. The platform allows for the execution of user-defined reaction protocols within an array of storage chambers by consecutively merging programmable sequences of pL-volume droplets containing specified reagents. The precision in formulating solutions with small differences in concentration is perfectly suited for the accurate estimation of kinetic parameters. The utility of our platform is showcased through the performance of enzymatic kinetic measurements of beta-galactosidase and horseradish peroxidase with fluorogenic substrates. The presented platform provides for a range of automated manipulations and paves the way for a more diverse range of droplet-based biological experiments.

Automated summary

Droplet-based microfluidic systems have become popular in high throughput screening due to their flexibility, high-throughput capabilities, and efficient processing of small volumes. However, the challenges of performing customized operations on user-defined droplets have limited their use in complex screening applications. To address this, the integration of droplet- and valve-based microfluidic technologies provides a solution for controlling droplet manipulations while maintaining analytical throughput. This article presents a microfluidic platform that combines microvalve technology with droplet-based compartmentalization to achieve programmable fluid handling. The platform allows for the execution of user-defined reaction protocols within storage chambers by merging droplets containing specified reagents.