RESEARCH
Research review
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#1 Artificial objects and Natural products
> In the nature, molecules assemble by themselves and produce a minute nanostructure
under a low energy (low temperature, ordinary pressure).
> Microfabrication can create controlled microstructures.
> It is difficult to create natural structures under a nanoscale (missing area).
> We construct a structure controlled sizes from an angstrom to a meter scale, embedding
the nano scale structure produced by living things in artifacts.
#2 Artificial cell membranes (Lipid Bilayer) and membrane proteins
> Phospholipid molecules spontaneously form bilayer lipid membranes by
Hydrophobic and Hydrophilic Interaction in the water.
> Membrane proteins perform various functions in the cell membranes.
> Some membrane protein transport substances, energy and information between
outside and inside the cells.
> Membrane proteins perform as the transporter of ions and medicines, the receptor
of odorant and light, and the perception of mechanical stress.
#3 MEMS(MicroElectroMechanicalSystem) and membrane proteins
In our laboratory,
> We manufacture the platform for preparing artificial cell membranes by using a micro electro
mechanical system (MEMS).
> System construction for measuring the signals of membrane protein reconstituted in lipid
bilayer membranes.
> Application for biosensor, drug development and physicochemical study in controlled
nano-space.
> Utilizing molecules, proteins, DNA/RNA, lipid, cell, MEMS, microfluidics techniques
in our study.
#4 Micro-sized processing techniques (Lithography, Milling)
> Microfabrication by photolithography with plasma-etching and exposure‑masking.
> CAD/CAM micromilling for plastic fabrication.
> Manufacture of electrodes by metal deposition and confirmation of processing accuracy
by a measuring microscope.
#5 Droplet contact method: Formation of artificial cell membranes
> Applying of oil/lipid solution and buffer solution into a chamber, two lipid monolayers
are formed at the surface of aqueous droplets.
> Two droplet contact together and bilayer lipid membrane can be formed.
> Artificial cell membranes mimicking natural cell membranes are able to be formed by
various types of lipid molecules.
#6 A electrical single molecule detection by nanopore sensing
> Ions pass through a nanopore (≈1.4 nm in diameter) reconstituted in membranes
cause current increase.
> A translocating molecules through the pore show the blocking current.
> DNA unzipping make the blocking time longer.
> Application: drug testing, food inspection, and diagnostic pathology.
#7 The mimetic device of cellular information transfer mechanism
> DNA computing, which uses programmed DNA as logic elements, enables large-scale
parallel computation which is difficult for existing computers.
> Although this calculation requires longer detection time because of DNA amplification
by PCR and detection by fluorescence imaging, we implemented a reduction of detection
time by electrical detection using nanopore sensing.
> Nanopores enable the detection and translocation to next droplet at the single molecule level.
#8 Electrical molecular computing using nanopores
> Micro RNA(miRNA) is non-coding RNA, and it shows overexpress in cancer.
> The pattern recognition is necessary because expression pattern of miRNAs differ with
cancer types.
> This system can detect overexpress miRNAs in cancer specific by combination of DNA computing
and nanopore measurement.
> Application to simple diagnosis is expected.
#9 Characterization of various nanopores
> Nanopores used for sensing include various types of biological and solid nanopores.
> Characterization of these nanopores can find out optimal nanopore for sensing.
> By using nanopores with large diameter, it is possible to detect large molecules such as
diagnostic marker protein with high sensitivity.
#10 High-throughput measurement and preparation of droplets
> We achieved an artificial system generating two conductance states by embedding
a synthetic metal-organic porous (MOP) into lipid bilayers.
> Two-pore channels which are expressed ubiquitously in animals and plants control
the resting membrane potential by using dual channels.
> We demonstrated two distinct ion conductance states by embedding a single MOP
molecule with the geometry of an Archimedean cuboctahedron.
> The triangular and square apertures in the cuboctahedron work independently as
ion-transporting pathways.
#11 High-throughput measurement and preparation of droplets
> Antimicrobial peptides kill cell bacteria by forming nanopore into lipid bilayer.
> Pore-forming activity of the antimicrobial peptide is measured by electrical measurement.
> Current signals are classified by those of shapes, and we are enable to consider the
movement of peptides on membrane.
> This study contributes to clear the mechanism of antimicrobial peptides and to apply
them as antimicrobial drugs
#12 High-throughput measurement and preparation of droplets
> Molecular robots are composed of sensors, calculators, and actuators that are
all implemented in liposomes or hydrogels.
> Synthetic ion channels(nanopores) are potential candidates for the sensor sections
of molecular robots.
#13 High-throughput measurement and preparation of droplets
> The droplet contact method (DCM) has been reported to prepare stable planar bilayer
lipid membranes (pBLMs).
> In this study, we applied a DCM strategy to pBLM formation using microfluidic techniques
and attempted to form double-stacked pBLMs in micro-meter scale.
> First, microchannels with micro pillars were designed via hydrodynamic simulations to
form a five-layered flow with aqueous and lipid/oil solutions.
> Then, pBLMs were successfully formed by controlling the pumping pressure of the
solutions and allowing contact between the two lipid monolayers.
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