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Synchronized Reagent Delivery in Double Emulsions for Triggering Chemical Reactions and Gene Expression

preprint
submitted on 16.02.2021, 17:04 and posted on 17.02.2021, 13:10 by Ariane Stucki, Petra Jusková, Nicola Nuti, Steven Schmitt, Petra Dittrich
Microfluidic methods to form single emulsion and double emulsion (DE) droplets have greatly enhanced the toolbox for high throughput screening for cell or enzyme engineering and drug discovery. However, remaining challenges in the supply of reagents into these enclosed nanoliter compartments limit the applicability of droplet microfluidics. Here, we introduce a strategy for on-demand delivery of reactants in DEs. We use lipid vesicles as transport carriers, which are co-encapsulated in double emulsions and release their cargo upon addition of an external trigger, here the anionic surfactant SDS. The reagent present inside the lipid vesicles stays isolated from the remaining content of the DE vessel until SDS enters the DE lumen and solubilizes the lipid bilayer. We demonstrate the versatility of the method with two critical applications, chosen as representative assays for high throughput screening. First, we trigger enzymatic reactions after releasing a reactant and second, we encapsulate bacteria and induce gene expression at a delayed time. The presented technique is compatible with the high throughput analysis of individual DE droplets using conventional flow cytometry as well as with microfluidic time-resolved studies. The possibility of delaying and controlling reagent delivery in current high throughput compartmentalization systems will significantly extend their range of applications e.g. for directed evolution, and further improve their compatibility with biological systems.

History

Email Address of Submitting Author

petra.juskova@bsse.ethz.ch

Institution

ETH Zurich, Department of Biosystems Science and Engineering

Country

Switzerland

ORCID For Submitting Author

0000-0002-7009-4263

Declaration of Conflict of Interest

The authors have declared no competing interest.