Building Dynamic Cellular Machineries in Droplet-Based Artificial Cells with Single-DropletTracking and Analysis

Although the application of droplet microfluidics has grown exponentially in chemistry and biology over the past decades, robust universal platforms for the routine generation and comprehensive analysis of droplet-based artificial cells are still rare. Here we report using microfluidic droplets to reproduce a variety of types of cellular machinery in in-vitro artificial cells. In combination with a unique image-based analysis method, the system enables full automation in tracking single droplets with high accuracy, high throughput, and high sensitivity. These powerful performances allow broad applicability evident in three representative droplet-based analytical prototypes that we develop for: (i) droplet digital detection, (ii) in vitro transcription and translation kinetic measurements, and (iii) spatiotemporal dynamics of cell-cycle oscillations. The capacities of this platform to generate, incubate, track and analyze individual microdroplets via real-time long-term imaging unleash its great potentials...