Development of an automation technique for the establishment of functional lipid bilayer arrays

Abstract

In the present work, a technique for establishing multiple black lipid membranes (BLMs) in arrays of micro structured ethylene tetrafluoroethylene (ETFE) films, and supported by a micro porous material was developed. Rectangular 8 × 8 arrays with apertures having diameters of 301 ± 5 µm were fabricated in ETFE Teflon film by laser ablation using a carbon dioxide laser. Multiple lipid membranes could be formed across the micro structured 8 × 8 array ETFE partitions. Success rates for the establishment of cellulose-supported BLMs across the multiple aperture arrays were above 95%. However, the time course of the membrane thinning process was found to vary considerably between multiple aperture bilayer experiments. An airbrush partition pretreatment technique was developed to increase the reproducibility of the multiple lipid bilayers formation during the time course from the establishment of the lipid membranes to the formation of bilayers. The results showed that multiple lipid bilayers could be reproducible formed across the airbrush-pretreated 8 × 8 rectangular arrays. The ionophoric peptide valinomycin was incorporated into established membrane arrays, resulting in ionic currents that could be effectively blocked by tetraethylammonium. This shows that functional bimolecular lipid membranes were established, and furthermore outlines that the established lipid membrane arrays could host functional membrane-spanning molecules.

Tailoring Properties of Biocompatible PEG-DMA Hydrogels with UV Light

“…In the design of large scale biomimetic membranes [13][14][15] becomes important. In previous work [10,16] we characterized chemically activated and photoactivated PEG-DMA and PEG-DA hydrogels with respect to volumetric stability, water permeability, ionic permeability and porosity and showed successful hydrogel encapsulation of multiple lipid membranes formed across an aperture array.…”

Surface Modifications of Support Partitions for Stabilizing Biomimetic Membrane Arrays

“…Also, a high value for the membrane capacitance (C m ) is desirable in sensors based on detecting changes in membrane impedance. Generally, the formation, stability, and electrical properties of large stable membranes are critically dependent on the coupling between the membrane and its supporting scaffold [8,22,23]. In the development of biomimetic membranes it therefore becomes important to ensure good coupling/ adhesion at the interface between the biomimetic membrane and its surrounding scaffold.…”

“…Recently, multi-aperture BLM arrays have attracted interest as a method to scale up the effective area of biomimetic membranes [8]. Functional biomimetic membranes formed across multiple apertures requires a high degree of stability and production reproducibility and both parameters are strongly influenced by the partition material properties.…”

“…Functional biomimetic membranes formed across multiple apertures requires a high degree of stability and production reproducibility and both parameters are strongly influenced by the partition material properties. Previously we described the development of an automation technique for establishing BLMs in arrays and we showed that variations in the pretreatment of the ETFE partition was a major determining factor for the ability to reproducibly establish lipid bilayers [8]. The reproducibility could however be increased significantly by developing a controllable airbrush pretreatment technique.…”

“…HDMSO was chosen because of its ability to form hydrophobic layers on polymeric films by plasma polymerization [2,26], The olefins 1-decene and n-hexene were chosen as a mimic to traditional prepainting using a low molecular weight hydrocarbon [8].…”