Challenges and opportunities in achieving the full potential of droplet interface bilayers

Stephenson, Elanna B.; Korner, Jaime L.; Elvira, Katherine S.

doi:10.1038/s41557-022-00989-y

Cited by 11 publications

(7 citation statements)

References 95 publications

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“…The membrane tension can be estimated from the contact angles of the oil cap with the aqueous and agar phases ( see Supplementary Information ) and yields mN/m (obtained from 13 images on N=8 pseudo-vesicles), which is much larger than values usually reported in electroformed lipid vesicles 35 , 36 , mN/m. On the contrary, our membrane tension value compares well with those obtained in Droplet Interface Bilayers (DIBs) which are planar lipid bilayers obtained by putting in contact two aqueous droplets bathing in an oil+lipid mixture (see for instance 37 – 39 ). In DIBs, there is also water/oil interfaces surrounding the lipid bilayer.…”

Section: Membrane Characterisation and Functionalizationsupporting

confidence: 76%

A simple method to make, trap and deform a vesicle in a gel

Tapie

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Montel

et al. 2023

Sci Rep

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We present a simple method to produce giant lipid pseudo-vesicles (vesicles with an oily cap on the top), trapped in an agarose gel. The method can be implemented using only a regular micropipette and relies on the formation of a water/oil/water double droplet in liquid agarose. We characterize the produced vesicle with fluorescence imaging and establish the presence and integrity of the lipid bilayer by the successful insertion of $$\alpha $$ α -Hemolysin transmembrane proteins. Finally, we show that the vesicle can be easily mechanically deformed, non-intrusively, by indenting the surface of the gel.

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Section: Membrane Characterisation and Functionalizationsupporting

confidence: 76%

A simple method to make, trap and deform a vesicle in a gel

Tapie

Prevost

Montel

et al. 2023

Sci Rep

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“…DIBs have become the platform of choice for insertion of membrane proteins and ion channels into bilayers since they offer facile ways to form model membranes that allow for investigation of bilayer electrical and physical characteristics, , and droplets can be configured or perfused to deliver proteins or other solutes that interact with proteins. − The DIB model membrane is constructed by the juxtaposition of micrometer-size aqueous droplets, each immersed in an oil (e.g., liquid hydrocarbon) medium and each surrounded by a monolayer of amphiphile (e.g., phospholipid) assembled at the water–oil interface. When the two droplets physically adjoin, a lipid bilayer forms at the interdroplet region due to apposition of monolayers from each droplet. , A wide variety of uncharged and charged phospholipids have already been employed in the literature to form DIBs, either singly or in several combinations, and there is a growing interest in creating DIBs that better mimic the lipid composition of actual cellular membranes in order to host proteins and assay their function. But given that many important phospholipids have nonzero intrinsic curvature, it is important to recognize that the curvature stress existing in a DIB is not directly known.…”

Section: Introductionmentioning

confidence: 99%

The Role of Lipid Intrinsic Curvature in the Droplet Interface Bilayer

Gudyka,

Ceja-Vega,

Krmic

et al. 2024

Langmuir

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Model bilayers are constructed from lipids having different intrinsic curvatures using the droplet interface bilayer (DIB) method, and their static physicochemical properties are determined. Geometrical and tensiometric measurements are used to derive the free energy of formation (ΔF) of a two-droplet DIB relative to a pair of isolated aqueous droplets, each decorated with a phospholipid monolayer. The lipid molecules employed have different headgroup sizes but identical hydrophobic tail structure, and each is characterized by an intrinsic curvature value (c 0 ) that increases in absolute value with decreasing size of headgroup. Mixtures of lipids at different ratios were also investigated. The role of curvature stress on the values of ΔF of the respective lipid bilayers in these model membranes is discussed and is illuminated by the observation of a decrement in ΔF that scales as a near linear function of c 0 2 . Overall, the results reveal an association that should prove useful in studies of ion channels and other membrane proteins embedded in model droplet bilayer systems that will impact the understanding of protein function in cellular membranes composed of lipids of high and low curvature.

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“…To meet these demands, one feasible approach is to develop a high-throughput and low-sample-volume articial lipid bilayer formation system. 21 Conventionally, the "painting", "folding" or "vesicle" methods 22,23 are used for bilayer formation, which are well-established but challenging to integrate and scale up for high-throughput applications. Current studies make use of microchips to develop platforms for multichannel Black Lipid Membrane (BLM) formation.…”

Section: Introductionmentioning

confidence: 99%