Mechanical Actuation on Surface (MAOS) Microfluidics: Compress to Preparation for Next-Generation Sequencing

Abstract

We present Mechanical Actuation on Surface (MAOS), a programmable microfluidic platform that manipulates droplets via localized mechanical compression—eliminating the need for embedded electronics or fixed microchannel geometries. MAOS integrates essential fluidic operations—including droplet transport, magnetic bead-based purification, and thermal cycling—within a compact benchtop instrument and single-use cartridge. The system accommodates droplet volumes from nanoliters to microliters, enabling precise control over sequential biochemical processes. By studying the dynamic behavior of diverse fluids under compression, we identified the key physical variables—surface tension, contact angle, and viscosity—that dictate the onset of droplet motion. We observed sharp transitions in mobility around specific thresholds, and validated interfacial encapsulation as a general strategy to overcome resistive pinning. We validated MAOS by implementing a miniaturized next-generation sequencing (NGS) library preparation workflow. Magnetic bead-based cleanup demonstrated high-fidelity DNA recovery and fragment size selection comparable to manual methods, while PCR amplification was performed reliably in low-volume (5 µL) reactions with minimal evaporation. We then executed the full NGS library preparation workflow in a high-throughput format, processing eight libraries in parallel on a single disposable cartridge using as little as 10% of standard reagent volumes. Both short- and long-read sequencing outputs from MAOS-multiplexed libraries matched manual protocols across all key quality metrics. These results establish MAOS as a scalable and user-friendly alternative to conventional microfluidics, suitable for diverse applications in molecular biology, chemistry, and high-throughput workflows.