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FACS-Sortable Triple Emulsion Picoreactors for Screening Reactions in Biphasic Environments
Summary
Researchers developed FACS-sortable triple emulsion picoreactors with a fluorocarbon shell and biphasic octanol-in-water core for high-throughput screening of biphasic chemical reactions, optimising surfactants to produce stable emulsions compatible with multi-hour assays and screening millions of picoliter reactors using fluorescence-activated cell sorting.
ABSTRACT Biphasic environments can enable successful chemical reactions where any single solvent results in poor substrate solubility or poor catalyst reactivity. For screening biphasic reactions at high-throughput, a platform based on microfluidic double emulsions could use widely available FACS (Fluorescence Activated Cell Sorting) machines to screen millions of picoliter reactors in a few hours. However, encapsulating biphasic reactions within double emulsions to form FACS-sortable droplet picoreactors requires optimized solvent phases and surfactants to produce triple emulsion droplets that are stable over multi-hour assays and compatible with desired reaction conditions. This work demonstrates such FACS-sortable triple emulsion picoreactors with a fluorocarbon shell and biphasic octanol-in-water core. First, surfactants were screened to stabilize octanol-in-water emulsions for the picoreactor core. With these optimized conditions, stable triple emulsion picoreactors were produced (>70% of droplets survived to 24 hours), and the ability to produce protein in the biphasic core was demonstrated via cell-free protein synthesis. Finally, triple emulsion picoreactors were sorted based on fluorescence using commercial FACS sorters at >100 Hz with 75-80% of droplets recovered. These triple emulsion picoreactors have potential for future screening bead-encoded catalyst libraries, including enzymes such as lipases for biofuel production.
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