Dataset Information

Microdroplet Actuation via Light Line Optoelectrowetting (LL-OEW).

ABSTRACT: Meanwhile, electrowetting-on-dielectric (EWOD) is a well-known phenomenon, even often exploited in active micro-optics to change the curvature of microdroplet lenses or in analytical chemistry with digital microfluidics (DMF, lab on a chip 2.0) to move/actuate microdroplets. Optoelectrowetting (OEW) can bring more flexibility to DMF because in OEW, the operating point of the lab chip is locally controlled by a beam of light, usually impinging onto the chip perpendicularly. As opposed to pure EWOD, for OEW, none of the electrodes has to be structured, which makes the chip design and production technology simpler; the path of any actuated droplet is determined by the movement of the light spot. However, for applications in analytical chemistry, it would be helpful if the space both below as well as that above the lab chip were not obstructed by any optical components and light sources. Here, we report on the possibility to actuate droplets by laser light beams, which traverse the setup parallel to the chip surface and inside the OEW layer sequence. Since microdroplets are grabbed by this surface-parallel, nondiverging, and nonexpanded light beam, we call this principle "light line OEW" (LL-OEW).

SUBMITTER: Doering C

PROVIDER: S-EPMC8718280 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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Publications

Microdroplet Actuation via Light Line Optoelectrowetting (LL-OEW).

Doering Christoph C Strassner Johannes J Fouckhardt Henning H

International journal of analytical chemistry 20211223

Meanwhile, electrowetting-on-dielectric (EWOD) is a well-known phenomenon, even often exploited in active micro-optics to change the curvature of microdroplet lenses or in analytical chemistry with digital microfluidics (DMF, lab on a chip 2.0) to move/actuate microdroplets. Optoelectrowetting (OEW) can bring more flexibility to DMF because in OEW, the operating point of the lab chip is locally controlled by a beam of light, usually impinging onto the chip perpendicularly. As opposed to pure EWO ...[more]

PMID: 34976066

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Project description:Electroactive organisms contribute to metal cycling, pollutant removal, and other redox-driven environmental processes via extracellular electron transfer (EET). Unfortunately, developing genotype-phenotype relationships for electroactive organisms is challenging because EET is necessarily removed from the cell of origin. Microdroplet emulsions, which encapsulate individual cells in aqueous droplets, have been used to study a variety of extracellular phenotypes but have not been applied to investigate EET. Here, we describe the development of a microdroplet emulsion system to sort and enrich EET-capable organisms from complex populations. We validated our system using the model electrogen Shewanella oneidensis and described the tooling of a benchtop microfluidic system for oxygen-limited conditions. We demonstrated the enrichment of strains exhibiting electroactive phenotypes from mixed wild-type and EET-deficient populations. As a proof-of-concept application, we collected samples from iron sedimentation in Town Lake (Austin, TX) and subjected them to microdroplet enrichment. We measured an increase in electroactive organisms in the sorted population that was distinct compared to a population growing in bulk culture with Fe(III) as the sole electron acceptor. Finally, two bacterial species not previously shown to be EET-capable, Cronobacter sakazakii and Vagococcus fessus, were further cultured and characterized for electroactivity. Our results demonstrate the utility of microdroplet emulsions for isolating and identifying EET-capable bacteria.IMPORTANCEThis work outlines a new high-throughput method for identifying electroactive bacteria from mixed populations. Electroactive bacteria play key roles in iron trafficking, soil remediation, and pollutant degradation. Many existing methods for identifying electroactive bacteria are coupled to microbial growth and fitness-as a result, the contributions from weak or poor-growing electrogens are often muted. However, extracellular electron transfer (EET) has historically been difficult to study in high-throughput in a mixed population since extracellular reduction is challenging to trace back to the parent cell and there are no suitable fluorescent readouts for EET. Our method circumvents these challenges by utilizing an aqueous microdroplet emulsion wherein a single cell is statistically isolated in a pico- to nano-liter-sized droplet. Then, via fluorescence obtained from copper reduction, the mixed population can be fluorescently sorted and gated by performance. Utilizing our technique, we characterize two previously unrecognized weak electrogens Vagococcus fessus and Cronobacter sakazakii.

Dataset Information

Microdroplet Actuation via Light Line Optoelectrowetting (LL-OEW).

Publications

Microdroplet Actuation via Light Line Optoelectrowetting (LL-OEW).

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