Passive and parallel microfluidic formation of droplet interface bilayers (DIBs) for measurement of leakage of small molecules through artificial phospholipid membranes

Czekalska, Magdalena A.; Kamiński, Tomasz S.; Makuch, Karol; Garstecki, Piotr

doi:10.1016/j.snb.2019.01.143

Cited by 20 publications

(16 citation statements)

References 47 publications

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“… 21 DIB formation automation using droplet microfluidics has yielded several promising systems for imaging molecular transport, wherein droplet pairs are precisely positioned and monitored over time. 22 – 24 These systems have limited throughput, however, and semi-stochastic droplet-droplet contact further complicates accurate transport measurement. The present device forms hundreds of DIBs per minute in a controlled and continuous process, minimizing variation and greatly enhancing statistical power.…”

Section: Resultsmentioning

confidence: 99%

Chiral lipid bilayers are enantioselectively permeable

Cochrane

Jones

et al. 2021

Nat. Chem.

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Homochiral membrane bilayers organize biological functions in all domains of life. The membrane’s permeability–its key property–correlates with a molecule’s lipophilicity, but the role of the membrane’s rich and uniform stereochemistry as a permeability determinant is largely ignored in empirical and computational measurements. Here, we describe a new approach to measuring permeation using continuously generated microfluidic droplet interface bilayers (DIBs, 480/min) and benchmark this system by monitoring fluorescent dye DIB permeation over time. Permeation of non-fluorescent, alkyne-labeled molecules was measured using a fluorogenic click reaction. DIB transport measurements revealed enantioselective permeation of alkyne-labeled amino acids (Ala, Val, Phe, Pro) and dipeptides through a chiral phospholipid bilayer; the biological L enantiomers permeated faster than D (1.2–6-fold; Ala–Pro). Enantioselective permeation both poses a potentially unanticipated criterion for drug design and offers a kinetic mechanism for the abiotic emergence of homochirality via chiral transfer between sugars, amino acids, and lipids.

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Section: Resultsmentioning

confidence: 99%

Chiral lipid bilayers are enantioselectively permeable

Cochrane

Jones

et al. 2021

Nat. Chem.

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“…The proposed microfluidic architecture offers several advantages compared to others based on serial (Schlicht and Zagnoni 2015;Nguyen et al 2016) or parallel (Czekalska et al 2019) DIBs designs. These are: its scalability, the way DIBs can be formed within a few seconds of each other and the robustness of achieving droplet pairing in AB configuration and in parallel fashion.…”

Section: Discussionmentioning

confidence: 99%

“…The combination of droplet microfluidics with dropletinterface-bilayer (DIB) protocols (Schlicht and Zagnoni 2015;Trantidou et al 2018) offers potentially large-scale solutions to overcome the limitations associated with the current live cell-based techniques, generating artificial cell membrane structures within which ion channels can be studied in simplified synthetic microenvironments (Bayley et al 2008). Of importance when investigating ion channel functions in microfluidic DIBs is (i) the control over droplet pair matching and positioning, (ii) guaranteeing automation of the microfluidic protocols and (iii) achieving large data throughput (Nguyen et al 2016;Czekalska et al 2019). This approach is therefore promising for studying intracellular ion channels, complementing life-cell electrophysiology data in situations where ion channel location or overexpression in live-cell is problematic.…”

Section: Introductionmentioning

confidence: 99%

Functionalisation of human chloride intracellular ion channels in microfluidic droplet-interface-bilayers

Zhang

Bracken

Woolhead

et al. 2020

Biosensors and Bioelectronics

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Profiling ion flux through human intracellular chloride ion channels using live-cell based techniques, such as patch-clamp electrophysiology, is laborious and time-consuming. The integration of scalable microfluidic systems with automatable protocols based on droplet-interface-bilayers (DIBs) within which ion channels are incorporated circumvents several limitations associated with live-cell measurements and facilitates testing in controllable in vitro conditions. Here, we have designed and tested novel microfluidic layouts for the formation of arrays of DIBs in parallel and developed the first example of a miniaturised, DIB-based, fluorescence assays for Clfluxing, allowing the investigation of the functional properties of the human chloride intracellular ion channel 1 (CLIC1). The microfluidic protocols relied on passive geometries for droplet pairing and DIB formation. Using recombinantly expressed CLIC1, we identified the best conditions to maximise protein integration into a lipid bilayer and the oligomerisation of the protein into functional ion channels. Finally, CLIC1 ion channel functionality was assessed relative to α-Haemolysin into microfluidic DIBs using the same Clfluxing assay.

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“…For example, a microfluidic platform containing a set of T-junctions was reported for droplet formation from multiple samples, with generated droplets isolated in a trap array. 36 Although this device uses a simple strategy to form and store a droplet array, the trapped droplets cannot be retrieved for further operations. In a related system, metered volumes of up to eight different samples were sequestered within individual traps or combined within a single trap, with the recovery of selected droplets from a large-scale array achieved by hydrodynamic flow with on-chip multiplexed valving.…”

Section: Introductionmentioning

confidence: 99%

A programmable microfluidic platform for multisample injection, discretization, and droplet manipulation

et al. 2020

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A programmable microfluidic platform enabling on-demand sampling, compartmentalization, and manipulation of multiple aqueous volumes is presented. The system provides random-access actuation of a microtrap array supporting selective discretization of picoliter volumes from multiple sample inputs. The platform comprises two interconnected chips, with parallel T-junctions and multiplexed microvalves within one chip enabling programmable injection of aqueous sample plugs, and nanoliter volumes transferred to a second microtrap array chip in which the plugs are actively discretized into picoliter droplets within a static array of membrane displacement actuators. The system employs two different multiplexer designs that reduce the number of input signals required for both sample injection and discretization. This versatile droplet-based technology offers flexible sample workflows and functionalities for the formation and manipulation of heterogeneous picoliter droplets, with particular utility for applications in biochemical synthesis and cell-based assays requiring flexible and programmable operation of parallel and multistep droplet processes. The platform is used here for the selective encapsulation of differentially labeled cells within a discrete droplet array.

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Passive and parallel microfluidic formation of droplet interface bilayers (DIBs) for measurement of leakage of small molecules through artificial phospholipid membranes

Cited by 20 publications

References 47 publications

Chiral lipid bilayers are enantioselectively permeable

Chiral lipid bilayers are enantioselectively permeable

Functionalisation of human chloride intracellular ion channels in microfluidic droplet-interface-bilayers

A programmable microfluidic platform for multisample injection, discretization, and droplet manipulation

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