High-throughput double emulsion-based microfluidic production of hydrogel microspheres with tunable chemical functionalities toward biomolecular conjugation

We have developed a novel workflow (sdDE-FACS, s̲ingle d̲roplet D̲ouble E̲mulsion FACS) that allows robust production, screening, and sorting of single double emulsion droplets with complete nucleic acid recovery.

Download Full-text

Porosity-Tuned Chitosan–Polyacrylamide Hydrogel Microspheres for Improved Protein Conjugation

Biomacromolecules ◽

10.1021/acs.biomac.6b00582 ◽

2016 ◽

Vol 17 (7) ◽

pp. 2427-2436 ◽

Cited By ~ 13

Author(s):

Sukwon Jung ◽

John H. Abel ◽

Jesse L. Starger ◽

Hyunmin Yi

Keyword(s):

Polyacrylamide Hydrogel ◽

Protein Conjugation ◽

Hydrogel Microspheres

Download Full-text

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

10.26434/chemrxiv.14039825.v1 ◽

2021 ◽

Author(s):

Ariane Stucki ◽

Petra Jusková ◽

Nicola Nuti ◽

Steven Schmitt ◽

Petra Dittrich

Keyword(s):

Gene Expression ◽

High Throughput ◽

High Throughput Screening ◽

Double Emulsion ◽

Lipid Vesicles ◽

Enzyme Engineering ◽

Droplet Microfluidics ◽

Enzymatic Reactions ◽

High Throughput Analysis ◽

Double Emulsions

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.

Download Full-text

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

10.26434/chemrxiv.14039825 ◽

2021 ◽

Author(s):

Ariane Stucki ◽

Petra Jusková ◽

Nicola Nuti ◽

Steven Schmitt ◽

Petra Dittrich

Keyword(s):

Gene Expression ◽

High Throughput ◽

High Throughput Screening ◽

Double Emulsion ◽

Lipid Vesicles ◽

Enzyme Engineering ◽

Droplet Microfluidics ◽

Enzymatic Reactions ◽

High Throughput Analysis ◽

Double Emulsions

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.

Download Full-text