Handling of artificial membranes using electrowetting-actuated droplets on a microfluidic device combined with integrated pA-measurements

Martel, Anne; Cross, Benjamin

doi:10.1063/1.3665719

Cited by 8 publications

(9 citation statements)

References 27 publications

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“…DIB formation is achieved when two psW/O droplets come into contact and the lipid molecules at each droplet interface interact spontaneously self-assembling into a lipid bilayer. Initially, DIB based assays have been developed by manually bringing droplets into contact but, in recent years, more efforts have been devoted to establishing similar assays using miniaturised systems that relied on micromanipulators, microgeometries and dielectrophoresis 9 10 11 12 13 14 15 16 or by interfacing psW/O droplets with agarose gel layers 17 18 . However, these approaches present limitations due to either a lower throughput than automated patch-clamp systems or necessitate procedures that require manual intervention.…”

mentioning

confidence: 99%

Droplet-interface-bilayer assays in microfluidic passive networks

Schlicht

Zagnoni

2015

Sci Rep

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Basic biophysical studies and pharmacological processes can be investigated by mimicking the intracellular and extracellular environments across an artificial cell membrane construct. The ability to reproduce in vitro simplified scenarios found in live cell membranes in an automated manner has great potential for a variety of synthetic biology and compound screening applications. Here, we present a fully integrated microfluidic system for the production of artificial lipid bilayers based on the miniaturisation of droplet-interface-bilayer (DIB) techniques. The platform uses a microfluidic design that enables the controlled positioning and storage of phospholipid-stabilized water-in-oil droplets, leading successfully to the scalable and automated formation of arrays of DIBs to mimic cell membrane processes. To ensure robustness of operation, we have investigated how lipid concentration, immiscible phase flow velocities and the device geometrical parameters affect the system performance. Finally, we produced proof-of-concept data showing that diffusive transport of molecules and ions across on-chip DIBs can be studied and quantified using fluorescence-based assays.

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mentioning

confidence: 99%

Droplet-interface-bilayer assays in microfluidic passive networks

Schlicht

Zagnoni

2015

Sci Rep

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“…Reducing the membrane surface tension enhances its favorability in the system leading to a relaxation or expansion in its area. This phenomena is recently used as the driving force for multiple droplet on a microchip manipulations [ 174 , 175 ] as well as pore gating through membrane tension alterations [ 173 ]. However, this expansion is not always possible given boundary conditions and constraints on the model membrane.…”

Section: Electrophysiological Methods For Characterizing Lipid Membranesmentioning

confidence: 99%

Characterizing the Structure and Interactions of Model Lipid Membranes Using Electrophysiology

El-Beyrouthy

Freeman

2021

Membranes

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The cell membrane is a protective barrier whose configuration determines the exchange both between intracellular and extracellular regions and within the cell itself. Consequently, characterizing membrane properties and interactions is essential for advancements in topics such as limiting nanoparticle cytotoxicity. Characterization is often accomplished by recreating model membranes that approximate the structure of cellular membranes in a controlled environment, formed using self-assembly principles. The selected method for membrane creation influences the properties of the membrane assembly, including their response to electric fields used for characterizing transmembrane exchanges. When these self-assembled model membranes are combined with electrophysiology, it is possible to exploit their non-physiological mechanics to enable additional measurements of membrane interactions and phenomena. This review describes several common model membranes including liposomes, pore-spanning membranes, solid supported membranes, and emulsion-based membranes, emphasizing their varying structure due to the selected mode of production. Next, electrophysiology techniques that exploit these structures are discussed, including conductance measurements, electrowetting and electrocompression analysis, and electroimpedance spectroscopy. The focus of this review is linking each membrane assembly technique to the properties of the resulting membrane, discussing how these properties enable alternative electrophysiological approaches to measuring membrane characteristics and interactions.

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“…38,39 Alternatively, a single aqueous volume can be divided into multiple compartments and then attached together using a exible substrate. 28,40 Microuidic DIBs have also been fabricated using dielectrophoresis, 41 electrowetting on dielectric (EWOD), 42,43 thin tubes, 44,45 ow focusing, 46 or 3D printing. 47 Recently, it has become apparent that DIBs offer distinct advantages for dynamic membrane characterization.…”

Section: Introductionmentioning

confidence: 99%

Dynamic morphologies of microscale droplet interface bilayers

et al. 2014

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Droplet interface bilayers (DIBs) are a powerful platform for studying the dynamics of synthetic cellular membranes; however, very little has been done to exploit the unique dynamical features of DIBs. Here, we generate microscale droplet interface bilayers (μDIBs) by bringing together femtoliter-volume water droplets in a microfluidic oil channel, and characterize morphological changes of the μDIBs as the droplets shrink due to evaporation. By varying the initial conditions of the system, we identify three distinct classes of dynamic morphology. (1) Buckling and fission: when forming μDIBs using the lipid-out method (lipids in oil phase), lipids in the shrinking monolayers continually pair together and slide into the bilayer to conserve their mass. As the bilayer continues to grow, it becomes confined, buckles, and eventually fissions one or more vesicles. (2) Uniform shrinking: when using the lipid-in method (lipids in water phase) to form μDIBs, lipids uniformly transfer from the monolayers and bilayer into vesicles contained inside the water droplets. (3) Stretching and unzipping: finally, when the droplets are pinned to the wall(s) of the microfluidic channel, the droplets become stretched during evaporation, culminating in the unzipping of the bilayer and droplet separation. These findings offer a better understanding of the dynamics of coupled lipid interfaces.

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Handling of artificial membranes using electrowetting-actuated droplets on a microfluidic device combined with integrated pA-measurements

Cited by 8 publications

References 27 publications

Droplet-interface-bilayer assays in microfluidic passive networks

Droplet-interface-bilayer assays in microfluidic passive networks

Characterizing the Structure and Interactions of Model Lipid Membranes Using Electrophysiology

Dynamic morphologies of microscale droplet interface bilayers

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