Impacts of electrical parameters on the electroformation of giant vesicles on ITO glass chips

Li, Wen-Man; Wang, Qiong; Ye, Zhong; Wang, Wangang; Cao, Yi; Hu, Ning; Luo, Haojun; Liao, Yanjian; Yang, Jun

doi:10.1016/j.colsurfb.2015.11.020

Cited by 21 publications

(19 citation statements)

References 30 publications

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“…GUVs in the range of 50–100 μm or even larger can be formed in water or in sucrose solution by applying an AC voltage around 2–3 V pp for 2 h at 10 Hz (Herold et al, 2012 ). Optimization of the frequency protocol can lead to an increased GUV yield as shown by Li et al for GUV formation in pure water (Li et al, 2016 ). Figure 5 shows GUVs prepared in deionized water and an example of a supergiant vesicle of 220 μm diameter prepared in 300 mM sucrose.…”

Section: Electroformationmentioning

confidence: 98%

Production of Isolated Giant Unilamellar Vesicles under High Salt Concentrations

et al. 2017

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The cell membrane forms a dynamic and complex barrier between the living cell and its environment. However, its in vivo studies are difficult because it consists of a high variety of lipids and proteins and is continuously reorganized by the cell. Therefore, membrane model systems with precisely controlled composition are used to investigate fundamental interactions of membrane components under well-defined conditions. Giant unilamellar vesicles (GUVs) offer a powerful model system for the cell membrane, but many previous studies have been performed in unphysiologically low ionic strength solutions which might lead to altered membrane properties, protein stability and lipid-protein interaction. In the present work, we give an overview of the existing methods for GUV production and present our efforts on forming single, free floating vesicles up to several tens of μm in diameter and at high yield in various buffer solutions with physiological ionic strength and pH.

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Section: Electroformationmentioning

confidence: 98%

Production of Isolated Giant Unilamellar Vesicles under High Salt Concentrations

et al. 2017

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“…The inlet and outlet were left open to avoid bubble formation when the experimental setup was completed. The miniaturized reactor was fabricated based on a well-established soft lithography technique following reference [ 16 ]. A PMMA mold was made first by a laser marking machine.…”

Section: Methodsmentioning

confidence: 99%

Frequency-Dependent Electroformation of Giant Unilamellar Vesicles in 3D and 2D Microelectrode Systems

et al. 2017

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A giant unilamellar vesicle (GUV), with similar properties to cellular membrane, has been widely studied. Electroformation with its simplicity and accessibility has become the most common method for GUV production. In this work, GUV electroformation in devices with traditional 3D and new 2D electrode structures were studied with respect to the applied electric field. An optimal frequency (10 kHz in the 3D and 1 kHz in the 2D systems) was found in each system. A positive correlation was found between GUV formation and applied voltage in the 3D electrode system from 1 to 10 V. In the 2D electrode system, the yield of the generated GUV increased first but decreased later as voltage increased. These phenomena were further confirmed by numerically calculating the load that the lipid film experienced from the generated electroosmotic flow (EOF). The discrepancy between the experimental and numerical results of the 3D electrode system may be because the parameters that were adopted in the simulations are quite different from those of the lipid film in experiments. The lipid film was not involved in the simulation of the 2D system, and the numerical results matched well with the experiments.

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“…Although the exact mechanism of the method is not yet completely understood, it is believed that the electric field affects lipid swelling through direct electrostatic interactions, redistribution of counterions, changes in membrane surface and line tension, and electroosmotic flow effects [32]. More detailed theoretical discussions on the electroformation mechanism can be found elsewhere [33][34][35]; here we focus mainly on exploring the optimal parameters and artifacts appearing when performing experimental work using this method.…”

Section: Classic Electroformation Protocolmentioning

confidence: 99%

“…Most often, a separation value from this interval is used without special explanation, and the voltage is modified accordingly. In combination with ITO electrodes, the separation is achieved through the use of spacers usually made from silicon (PDMS) [2,31,34,37,[50][51][52] or Teflon (polytetrafluoroethylene) [28,38,41,83]. Another alternative is specifying the electric field strength instead of the voltage from the beginning [4,28,51].…”

Section: Electrical Parametersmentioning

confidence: 99%

Giant Unilamellar Vesicle Electroformation: What to Use, What to Avoid, and How to Quantify the Results

et al. 2021

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Since its inception more than thirty years ago, electroformation has become the most commonly used method for growing giant unilamellar vesicles (GUVs). Although the method seems quite straightforward at first, researchers must consider the interplay of a large number of parameters, different lipid compositions, and internal solutions in order to avoid artifactual results or reproducibility problems. These issues motivated us to write a short review of the most recent methodological developments and possible pitfalls. Additionally, since traditional manual analysis can lead to biased results, we have included a discussion on methods for automatic analysis of GUVs. Finally, we discuss possible improvements in the preparation of GUVs containing high cholesterol contents in order to avoid the formation of artifactual cholesterol crystals. We intend this review to be a reference for those trying to decide what parameters to use as well as an overview providing insight into problems not yet addressed or solved.

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Impacts of electrical parameters on the electroformation of giant vesicles on ITO glass chips

Cited by 21 publications

References 30 publications

Production of Isolated Giant Unilamellar Vesicles under High Salt Concentrations

Production of Isolated Giant Unilamellar Vesicles under High Salt Concentrations

Frequency-Dependent Electroformation of Giant Unilamellar Vesicles in 3D and 2D Microelectrode Systems

Giant Unilamellar Vesicle Electroformation: What to Use, What to Avoid, and How to Quantify the Results

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