The integrated droplet platform combines curved microstructures that allow high-efficiency (99.9%) reflow of droplets and a droplet cleaving that automatically synchronizes paired droplets enabling high-efficiency (99.9%) downstream merging.

Since being invented, droplet microfluidic technologies have been proven to be perfect tools for high-throughput chemical and biological functional screening applications, and they have been heavily studied and improved through the past two decades. Each droplet can be used as one single bioreactor to compartmentalize a big material or biological population, so millions of droplets can be individually screened based on demand, while the sorting function could extract the droplets of interest to a separate pool from the main droplet library. In this paper, we reviewed droplet detection and active sorting methods that are currently still being widely used for high-through screening applications in microfluidic systems, including the latest updates regarding each technology. We analyze and summarize the merits and drawbacks of each presented technology and conclude, with our perspectives, on future direction of development.

Droplet microfluidic systems have been widely deployed to interrogate biological and chemical systems. The major limitations of these systems are the relatively high error rates from critical droplet manipulation functions. To address these limitations, we describe the development of FIDELITY (Flotation and Interdigitated electrode forces on Droplets to Enable Lasting system IntegriTY), a highly sensitive and accurate size-based droplet bandpass filter that leverages the natural buoyancy of aqueous droplets and highly localized dielectrophoretic force generated by interdigitated electrode arrays. Droplet manipulation accuracies greater than 99% were achieved at a throughput of up to 100 droplets/s and separation of droplets that differed in diameter by only 6 μm was demonstrated. Last, the utility of FIDELITY was demonstrated in a droplet size quality control application and also in a droplet-based in vitro transcription/translation workflow. We anticipate FIDELITY to be integrated into a broad range of droplet microfluidic configurations to achieve exceptional operational accuracy.

Precise manipulation of cells within water-in-oil emulsion droplets has the potential to vastly expand the type of cellular assays that can be conducted in droplet-based microfluidics systems. However, achieving such manipulation remains challenging. Here, we present an in-droplet label-free cell separation technology by utilizing different dielectrophoretic responses of two different cell types. Two pairs of angled planar electrodes were utilized to generate positive or negative dielectrophoretic force acting on each cell type, which results in selective in-droplet movement of only one specific cell type at a time. A downstream asymmetric Y-shaped microfluidic junction splits the mother droplet into two daughter droplets, each of which contains only one cell type. The capability of this platform was successfully demonstrated by conducting in-droplet separation from a mixture of Salmonella cells and macrophages, two cell types commonly used as a bacterial pathogenicity analysis model. This technology enable the precise manipulation of cells within droplets, which can be exploited as a critical function in implementing broader ranges of droplet-based microfluidics cellular assays.

High-throughput screening techniques are pivotal to unlocking the mysteries of biology. Yet, the promise of droplet microfluidics in enabling single-cell resolution, ultra-high-throughput screening remains largely unfulfilled. Droplet sorting errors caused by polydisperse droplet sizes that are often inevitable in multi-step assays have severely limited the effectiveness and utility of this technique, especially when screening large libraries. Even a relatively low 1% sorting error results in 10,000 false calls in a 1,000,000 droplet screen, imposing an unreasonably large burden on downstream validation. Here, we present NOVAsort (Next-generation Opto-Volume-based Accurate droplet sorter), a device capable of discerning droplets based on both size and fluorescence intensity. With a 1000- and 10,000-fold reduction in false positives and false negatives, respectively. NOVAsort addresses the challenges of conventional droplet sorting approaches and sets standards for accuracy and throughput in droplet microfluidic assays.