Biomedical subjects

Hitoshi Muguruma

Publications and source records attributed to Hitoshi Muguruma.

9 recordsLinked to original sources

PubMed2006

Adsorption of glucose oxidase onto plasma-polymerized film characterized by atomic force microscopy, quartz crystal microbalance, and electrochemical measurement.

Adsorption of glucose oxidase (GOD) onto plasma-polymerized thin films (PPF) with nanoscale thickness was characterized by atomic force microscopy (AFM), quartz crystal microbalance (QCM), and electrochemical measurements. The PPF surface is very flat (less than 1-nm roughness), and its properties (charge and wettability) can be easily changed while retaining the backbone structure. We focused on three types of surfaces: (1) the pristine surface of hexamethyldisiloxane (HMDS) PPF (hydrophobic and neutral surface), (2) an HMDS PPF surface with nitrogen-plasma treatment (hydrophilic and positive-charged surface), and (3) an HMDS PPF surface treated with oxygen plasma (hydrophilic and negative-charged surface). The AFM image showed that the GOD molecules were densely adsorbed onto surface 2 and that individual GOD molecules could be observed. The longer axis of GOD ellipsoid molecules were aligned parallel to the surface, called the "lying position", because of electrostatic association. On surface 1, clusters of GOD molecules did not completely cover the original PPF surface (surface coverage was ca. 60%). The 10-nm-size step height between the GOD clusters and the PPF surface suggests that the longer axes of individual GOD molecules were aligned perpendicular to the surface, called the "standing position". On surface 3, only a few of the GOD molecules were adsorbed because of electrostatic repulsion. These results indicate that the plasma polymerization process can facilitate enhancement or reduction of protein adsorption. The AFM images show a corresponding tendency with the QCM profiles. The QCM data indicate that the adsorption behavior obeys the Langmuir isotherm equation. The amperometric biosensor characteristics of the GOD-adsorbed PPF on a platinum electrode showed an increment in the current because of enzymatic reaction with glucose addition, indicating that enzyme activity was mostly retained in spite of irreversible adsorption.

Adsorption↗

PubMed2006

Conformational polymorphism and thermochemical analysis of 5,5' ''-bis[(2,2,5,5-tetramethyl-1-aza-2,5-disila-1-cyclopentyl)ethyl]-2,2':5',2' ':5' ',2' ''-quaterthiophene.

The titled compound exists as two polymorphic solid phases (denoted form-I and form-II). Form-I obtained by as-synthesized material is a more stable phase. Form-II is a less stable phase. Spontaneous solid-solid transformation from form-II to form-I is observed in the temperature range between room temperature and the melting point of form-I (Tm = 156.5 degrees C), and its activation energy is estimated to be 96 kJ mol-1 by Arrhenius plot. The solid-solute-solid transformation (recrystallization from solution) from form-II to form-I is also observed. In contrast, form-II is obtained only by a solid-melt-solid transformation from form-I. Therefore, the system of two polymorphs is monotropic. The solid-state NMR measurement shows that form-I has the molecular conformation of complete S-syn-anti-syn in the oligothiophene backbone, whereas form-II has that of S-all-anti. With the solution NMR data, the polymorphism could not be observed. Therefore, the polymorphs originate from the different molecular packing involving the conformational change of the molecule. This unique property is attributed to the extra bulky terminal groups of the compounds. However, despite the extra bulky terminal groups, the mentioned polymorphism is not observed in the titled compound analogue which has S-all-anti conformation (like form-II).

Journal Article↗

PubMed2006

Structure and biosensor characteristics of complex between glucose oxidase and plasma-polymerized nanothin film.

The structure and biosensor characteristics of complex between glucose oxidase (GOD) and plasma-polymerized nanothin film (PPF), in which the thickness is several nanometers, were investigated by atomic force microscopy (AFM) and electrochemical measurement. The GOD molecules were densely adsorbed onto the PPF surface treated by nitrogen plasma and the individual GOD molecules were observed. Subsequently, the GOD densely packed array on the PPF surface was subsequently treated by plasma polymerization (overcoating). AFM image showed that the thicker film gave the smoother surface, indicating that the GOD adsorbed on the surface was embedded more deeply in PPF. The amperometric biosensor characteristics of the GOD-PPF complex on a platinum electrode showed the current increment due to the enzymatic reaction with glucose addition, indicating that enzyme activity was retained although the enzyme has been exposed to the plasma gas related to diffusion of the substrate. This means that under mild exposure to organic plasma, the enzyme does not become seriously dysfunctional. Amperometric biosensor characteristics were strongly affected by monomer and thickness of PPF overcoating related with the diffusion of the substrate (glucose). Considering that the film deposition was performed using microfabrication-compatible organic plasma, our new method for protein architecture has a great potential of enabling high throughput production of bioelectronic devices.

Biosensing Techniques↗

PubMed2005

Electron transfer mediator micro-biosensor fabrication by organic plasma process.

We propose a new strategy for constructing a mediator-type biosensor as a Bio-MicroElectroMechanical Systems (BioMEMS) application. A vinylferrocene plasma-polymerized film (PPF) was deposited directly onto the surface of an electrode under dry conditions. The resulting redox film was extremely thin, adhered well onto a substrate (electrode), and had a highly crosslinked network structure. This technique, capable of polymeric deposition of any kind of monomer, can also serve the purpose of anti-fouling coating, or layer-to-layer interface creation. With a subsequent plasma process, additional polymeric layer of hydrophilic acetonitrile was superimposed onto the existing vinylferrocene-PPF surface to offer crucial features such that the wettability could be adjusted for a better electron transfer, and amino functional groups could be attached to immobilize a large amount of enzyme. Based upon this scheme, the device fabrication could be designed in a manner that the whole procedure was made up of dry wafer-handling processes, which is compatible with mass production. A prototype device was fabricated to have an array of needle-shaped amperometric micro-biosensors. The resultant thin polymer layer carried a large number of the mediator molecules, accomplishing a lower overpotential (+410 mV) and a rapid response time (<5s). Stressing the advantages of the plasma polymerization process together with some additional features accomplished in our device fabrication, we would discuss new possibilities in the field of BioMEMS.

Biosensing Techniques↗

PubMed2005

Nanothin ferrocene film plasma polymerized over physisorbed glucose oxidase: high-throughput fabrication of bioelectronic devices without chemical modifications.

We describe a method for creating a mediator-containing interface between an enzyme and an electrode, achieving simpler and more reliable immobilization of the enzyme with the enhanced detection sensitivity. A nanothin polymer film containing a redox mediator, made of dimethylaminomethylferrocene, was plasma-deposited directly onto a glucose oxidase-physisorbed electrode, with which a relevant bioelectrochemical signal was observed without prior or further chemical modification of the enzyme molecules. The results of the surface characterizations before and after the enzyme immobilization showed that this method gave control over the spatial orientation of single enzyme molecules in favor of efficient and reproducible signal generation. Considering that the film deposition was performed using microfabrication-compatible organic plasma, our new method has a great potential of enabling high-throughput production of bioelectronic devices without chemical modification steps.

Adsorption↗

PubMed2005

Characterization of diffusion-controlled mass transport through nanoporous and nanothin films plasma polymerized on a sputtered platinum electrode.

We demonstrate the diffusion mode of various redox chemical species through a plasma-polymerized nanothin coating with nanometer-sized pores on a sputtered platinum (Pt) electrode. In this work, hexamethyldisiloxane plasma-polymerized films (PPFs) were added onto the sputtered platinum film, both of which were sequentially deposited in the same vacuum chamber. Results of atomic force microscopy, X-ray photoelectron spectroscopy, and electrochemical studies showed that the PPF provided the platinum electrode with a coating with a complete surface coverage. Sub-nanometer-sized pores (less than 1 nm) responsible for a highly crosslinked polymer network in the PPF coatings offered diffusivity-controlled permeation of redox molecules (i.e., size-exclusivity) rather than solubility-controlled permeation (i.e., chemoselectivity). Consequently, variation of the plasma power could give control over the size of the nanometer-sized cavities.

Journal Article↗

PubMed2004

Organic plasma process for simple and substrate-independent surface modification of polymeric BioMEMS devices.

A polymeric bio micro electromechanical systems (BioMEMS) device was fabricated using organic plasma polymerization, by which the surface of a polymeric substrate could easily be modified through vapor-phase deposition of organic thin films. This technique, capable of polymeric deposition of any kind of monomer, can serve the purpose of anti-fouling coating, wettability control, or layer-to-layer interface creation, on the surface of any given chemically-inert polymeric substrate without involving cumbersome surface organic reactions. A prototype device was fabricated to have an array of electrochemical glucose biosensors with the three electrode configuration, each of which has a microfluidic channel (500 microm x 800 microm) for capillary-action-driven sample delivery and the concerned enzymatic reaction. Stressing the advantages of the plasma polymerization process using a polymeric substrate together with some additional features accomplished in our device fabrication, new possibilities in the field of polymeric BioMEMS are discussed.

Biosensing Techniques↗

PubMed2004

Amperometric glucose biosensor based on mediated electron transfer between immobilized glucose oxidase and plasma-polymerized thin film of dimethylaminomethylferrocene on sputtered gold electrode.

We propose an electron transfer-mediated amperometric enzyme biosensor based on plasma-polymerized thin film of dimethylaminomethylferrocene (DMAMF) on a sputtered gold electrode. The DMAMF plasma-polymerized film is deposited directly onto the surface of the electrode under dry conditions. The resulting thin film not only has redox sites but also is extremely thin (approximately 20 nm), adheres well onto the substrate (electrode), has a flat surface and a highly-crosslinked network structure, and is hydrophilic in nature. Glucose oxidase is densely immobilized onto the surface of DMAMF plasma-polymerized film on the gold electrode. From the electrochemical measurement, the biosensor can cover the wide range of glucose concentration (1.3 - 81 mM) at +350 mV of applied potential. The current response of the glucose biosensor was decreased by less than 5% in an aerobic solution as compared to that in an anaerobic solution. These show that the DMAMF plasma-polymerized films play a role as the electron transfer mediators between the reaction center of enzyme and the electrode.

Biosensing Techniques↗

PubMed2002

MALC, a novel microinjection method for loading of macromolecules into cultured neurons.

Microinjection is one of the most useful and important methods in cellular neurobiology. However, direct insertion and retraction of a capillary produce physical stresses inside cells that make it difficult to apply on small and fragile cells, especially the central nervous system (CNS) neurons. In this study, we developed a novel method called MALC (microinjection method assisted with laser activated chromophore) to avoid this disadvantage of the conventional method. In MALC, a capillary is just attached on cell surface, and combination of a newly developed photosensitizer with a laser pointing system makes the plasma membrane permeable to molecules. This novel method can expand the application of the microinjection method and realize molecular manipulations inside CNS neurons.

Animals↗