Efficient Lipid Bilayer Formation by Dipping Lipid-Loaded Microperforated Sheet in Aqueous Solution

This paper describes a method for a bilayer lipid membrane (BLM) formation using a perforated sheet along with an open chamber. Microscopic observation of the formed membrane showed a typical droplet interface bilayer. We proved that the formed membrane was a BLM based on electrical measurements of the membrane protein α-hemolysin, which produces nanopores in BLMs. Unlike the conventional approach for BLM formation based on the droplet contact method, this method provides aqueous surfaces with no organic solvent coating layer. Hence, this method is suitable for producing BLMs that facilitate the direct addition of chemicals into the aqueous phase.

The role of temperature in the formation of human–mimetic artificial cell membranes using droplet interface bilayers (DIBs)

A systematic study of the role of temperature in human–mimetic droplet interface bilayer (DIB) formation.

Microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks

In the field of bottom-up synthetic biology, lipid membranes are the scaffold to create minimal cells and mimic reactions and processes at or across the membrane. In this context, we employ here a versatile microfluidic platform that enables precise positioning of nanoliter droplets with user-specified lipid compositions and in a defined pattern. Adjacent droplets make contact and form a droplet interface bilayer to simulate cellular membranes. Translocation of molecules across membranes are tailored by the addition of alpha-hemolysin to selected droplets. Moreover, we developed a protocol to analyze the translocation of non-fluorescent molecules between droplets with mass spectrometry. Our method is capable of automated formation of one- and two-dimensional droplet networks, which we demonstrated by connecting droplets containing different compound and enzyme solutions to perform translocation experiments and a multistep enzymatic cascade reaction across the droplet network. Our platform opens doors for creating complex artificial systems for bottom-up synthetic biology.

Enhanced water permeability across a physiological droplet interface bilayer doped with fullerenes

We measure the water permeability across a physiological lipid bilayer produced by the droplet interface bilayer technique.

Membrane determinants for the passive translocation of analytes through droplet interface bilayers

Identifying droplet interface bilayer conditions reproducing the permeability of cell membranes to small molecules.

Photo-Triggered Soft Materials With Differentiated Diffusive Pathways

Controlled diffusive transport between regions within a compartmentalized structure is an essential feature of cellular-inspired materials. Using the droplet interface bilayer (DIB) technique, biomolecular soft materials can be constructed in an oil medium by connecting multiple lipid-coated microdroplets together through interfacial bilayers. While traditionally achieved through the incorporation of pore forming toxins (PFTs), signal propagation within DIB assemblies can be remotely controlled through the integration of photopolymerizable phospholipids (23:2 DiynePC) into the aqueous phase. Since such strategy allows for the formation of UV-C triggered pathways only between droplets both containing DiynePC, polymerizable phospholipids have shown an advantage of reducing undesired diffusion and forming conductive pathways. The partial polymerization of lipid bilayers formed through the DIB platform is still to this date underexplored in the literature. In a previous work, we have shown that the incorporation of 23:2 DiynePC into lipid bilayers allows for the creation of patterned conductive pathways in a 2D DIB structure. The properties of photosensitive bilayers were also investigated but not their channel activity. The functionalization of bilayers-based photosensitive structures through transmembrane channels remains an under-investigated mean of achieving further differentiated conductive channels. This work explores the reconstitution of several transmembrane channels such as alpha-hemolysin (αHL) and alamethicin (ALM) into partially polymerized lipid bilayers. We believe that the ability to incorporate transmembrane channels into photosensitive DIB soft structures allows for further differentiation of signal propagation pathways by including both edge-defect induced pores as well as more traditional and bio-derived transporters.