Abstract
We describe the design, fabrication and use of a dual-layered microfluidic device for ultrahigh-throughput droplet trapping, analysis, and recovery using droplet buoyancy. To demonstrate the utility of this device for digital quantification of analytes, we quantify the number of droplets, which contain a β-galactosidase-conjugated bead among more than 100,000 immobilized droplets. In addition, we demonstrate that this device can be used for droplet clustering and real-time analysis by clustering several droplets together into microwells and monitoring diffusion of fluorescein, a product of the enzymatic reaction of β-galactosidase and its fluorogenic substrate FDG, between droplets.
Highlights
High-throughput analysis has become an important tool in biology and medicine for elucidating complex biological mechanisms, screening for therapeutic agents, and early diagnosis of disease [1]
With the above-mentioned advances and challenges in mind, we have developed a simple and robust dual-layered Floating Droplet Array (FDA) device for ultrahigh-throughput droplet manipulation, analysis, and recovery (Figure 1)
Β-gal beads and 500 μM FDG in PBS were introduced into the microfluidic device via respective inlets at a flow rate of 0.5 μL/min, while the oil phase was injected at a flow rate of 15 μL/min
Summary
High-throughput analysis has become an important tool in biology and medicine for elucidating complex biological mechanisms, screening for therapeutic agents, and early diagnosis of disease [1]. Microfabricated devices have emerged as a powerful experimental platform for performing a diverse range of biological and chemical assays in a high-throughput manner [3,4] These technologies often permit high-throughput analysis of a complex sample by partitioning a bulk solution into many isolated pico to nanoliter-sized compartments, or microreactors. Since the droplets are trapped in a secondary layer above the main flow stream, we are able to achieve high density and efficiency of immobilization for real-time, ultrahigh-throughput droplet analysis. DDrroopplleettss aarree rreeccoovveerreedd bbyy fflliippppiinngg tthhee ddeevviiccee ssoo tthhaatt ddrroopplleettss ffllooaatt oouutt ooff tthhee wweellllss ((vv)) aanndd aarree sseenntt ffoorr ddoowwnnssttrreeaamm hhaannddlliinngg oonn-- oorr ooffff--cchhiipp ((vvii)). PDMS layers were bonded immediately following oxygen plasma treatment and stored overnight before use
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