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This study reports an efficient droplet interface bilayer (DIB) formation using an open chamber with an external separator that can divide an aqueous solution to two regions. This approach facilitates reagent-saving and simplification of the device fabrication. Amphiphobic modification of the separator surface enables us to handle the device easily without liquid spill, and assists bilayer lipid membrane...
MicroRNA, a short-length RNA known as a biomarker of cancers, was detected by magnet-attached spiral channel and electrical measurement. A designed oligo-DNA bound with magnetic bead was entrapped in spiral channel by magnetic field, and hybridized with microRNA. Thereafter, liposome were exported from the spiral channel by duplex-specific nuclease, and applied to electrical measurement. Finally,...
This paper highlights the behavior of microdroplets used for portable biosensors. Microdroplets have been applied for various Lab-on-a-chip applications, in which portability/wearable is becoming a trend word. However, the resonance (sloshing phenomena) of the microdroplets could be an issue. For example, bilayer lipid membrane, fragile characteristics due to its thickness of 5 nm, easily ruptures...
This paper proposes a self-assembly method to construct a droplet interface bilayer (DIB) network. The DIB network has been developed for biological logical circuits or chemical microreactors, using incorporated membrane proteins. In this work, the DIB network was constructed by using multi-geometric Droplet-Boxes (DBs), which captured and stabilized an aqueous droplet in organic solvent. The multi-geometry...
MicroRNA, a biomarker for various diseases, was diagnosed by electrical monitoring of its transit event through a biological nanopore. The target microRNA in a sample solution selectively forms a duplex with a complementary DNA specially designed, which makes the transit time to slow down. An artificial lipid bilayer device previously reported allowed to implement the nanopore platform just by sequential...
This report describes preliminary results of a reconstitution and function of membrane proteins (flippases) into asymmetric lipid giant liposomes (GLs) that were prepared by deforming a planar asymmetric bilayer using a pulsed jet flow. Frist, we confirmed the expression of flippases on the baculovirus membrane by a western blot analysis. Next, we successfully reconstituted flippases into the asymmetric...
This paper proposes a cell pairing system that is capable of defining the number and the position of trapped cells by mechanically sliding the parylene rail films (PRF). The device can control the area of trapping sites of cells by sliding PRF on a SU-8 comb layer. This mechanism allows us to control the number as well as the order of lined-up cells. We successfully demonstrated lining up of three...
This paper describes a batch release technique for monodisperse liposomes immobilized on a substrate (Fig. 1). A single short-time pulsed voltage to the substrate induced detachment of the arrayed liposomes previously developed. Simultaneous release was observed shortly after the electrical stimulation. The release technique produced monodisperse and solvent-free liposomes freely suspended on the...
This paper aimed to develop a pendant liposome system that allows the access to both internal and external solutions of the formed liposomes. The access to the internal solution was achieved by a through-hole fabricated on the substrate, which was designed underneath the pendant-like, dome-shaped liposome. The formation of a uniform-size liposome array was based on our previous works that applied...
This paper describes a contactless manipulation system for giant liposomes in an aqueous flow by negative dielectrophoresis (nDEP). The system consists of 2 pairs of comb electrodes that face each other across a perpendicular channel. This design allows an nDEP force to be focused at the center of the gap of the electrodes and the liposomes to align in lines without contacting either the electrodes...
This paper describes the preparation of a cell-sized vesicle containing small vesicles from a two-planar lipid bilayer in a triple-well device using a pulsed jet flow. First, the size distribution of the cell-sized vesicles is estimated using a double-well device wherein a separator having a hole diameter of 100, 200, or 500 μm is mounted between the wells. Moreover, the small vesicles are successfully...
This paper describes microfluidic logic gates which use DNA and biological nanopores. Single-stranded DNA (ssDNA) can pass through αHL, a biological nanopore, incorporated in bilayer lipid membranes (BLMs), whereas double-stranded DNA (dsDNA) cannot. In this study, these passing and non-passing phenomena were applied as the binary system and logic gates. Two types of ssDNA were used as inputs, while...
This paper describes a bilayer lipid membranes (BLMs) chip capable of solution exchange. In the previous report, we successfully reconstituted α-hemolysin (αHL) in BLMs using droplets contacting method with an array device. For drug screening, solution exchange is required to test the effect of inhibitors or promoters for proteins. In this study, we attempted to exchange the solution in a droplet...
In this paper we propose an efficient tweezing methodology for liposome using positive-dielectrophoresis (pDEP). These DEP-based tweezers consist of a round-shaped glass-capillary equipped with two thin Au film electrodes. The electrodes are fabricated using a double side shadow mask (DSSM) technique. This very simple process enables low-cost and reliable fabrication. Using the DEP-based tweezers,...
This work presents two important technologies integrated in our uniform-size liposome array platform (Fig. 1a) to realize an artificial cell array system. One is the effective encapsulation of small objects in the arrayed liposomes (Fig. 1b top), making use of the electrospray deposition technique also used for the lipid patterning. The target nanobeads were selectively patterned on the lipid and...
Giant unilamellar vesicles (GUVs) are biochemical compartments enclosed with phospholipid bilayers. GUVs are expected to be materials for construction and investigation of artificial cell models. There, fusion phenomena between GUVs and smaller vesicles (referred to as "asymmetric fusion" here) seem to have some potential for utilization of the transporters bringing reagents and membrane...
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