Measurement of a Bubble-Free Chemical Oscillator Using QCMs Treated with Self-Assembled Monolayers.

The oscillation of a 1,4-cyclohexanedione-bromate (CHD-bromate) system was investigated by using a quartz crystal microbalance (QCM) without or with a self-assembled monolayer (SAM), where gold and platinum were used for QCM electrodes and SAMs were composed of HS(CH2)11CH3, HS(CH2CH2O)5H, and HS(CH2CH2O)5CH3. The CHD-bromate system is well-known as a bubble-free oscillator and oscillates without or with a catalyst. In the CHD-bromate system without a catalyst, the oscillation of a resonant frequency shift (ΔF) of the QCM was observed in the Au-electrode QCMs without a SAM or with SAMs formed from HS(CH2)11CH3 and HS(CH2CH2O)5H. On the other hand, the HS(CH2CH2O)5CH3 SAM suppressed the ΔF oscillation. The results revealed that in the CHD-bromate system without a catalyst, hydrophobic CH3 terminal and helical conformation were important to prevent nonspecific adsorption of substances on a gold surface and its dissolution. In the CHD-bromate system with a catalyst (ferroin), the ΔF oscillation was observed in the Au-electrode QCM with the HS(CH2CH2O)5CH3 SAM. The results suggest evidence that the change of the solution viscosity and density led to the ΔF oscillation. These results for Au-electrode QCMs were also corroborated by those for Pt-electrode QCMs.

Temperature Dependence of the Rheology of Soft Matter on a MHz-oscillating Solid-liquid Interface.

The temperature dependence of the resonant length, molecular weight, and rheology (shear viscosity and shear modulus) of chemisorbed soft matter on a solid-liquid interface oscillating at a megahertz frequency was studied using a quartz crystal microbalance. As a form of chemisorbed soft matter, self-assembled monolayers (SAMs) formed from six types of mercapto oligo(ethylene oxide) methyl ethers were used. A systematic analysis using the Voigt model showed that the variation in effective hydrated thickness (sensed mass), which is related to the resonant length, was classified into three types based on the molecular weight. As a result, a 2.2-nm change in the resonant length occurred in the studied temperature range from 10 to 35℃. Moreover, the variation in the effective hydrated thickness was dependent on the shear viscosity and shear modulus of the SAMs. A further investigation revealed that the relationships η 1∝M n 0.13 and µ 1∝M n 0.30 could be estimated regardless of the temperature, where η 1 and µ 1 are the shear viscosity and shear modulus of the SAM, and M n is the molecular weight of mercapto oligo(ethylene oxide) methyl ether. As a result, we revealed that the experimental results followed the polymer formula irrespective of temperature.

Surface Modification of PDMS and Plastics with Zwitterionic Polymers.

Surface modification of PDMS, polycarbonate, and acrylic resin was examined using various methacryl polymers bearing sulfobetaine, phosphoryl choline, and oligoethylene glycol units. We have found that zwitterionic polymers are adsorbed on the PDMS surface treated with plasma. The surface of PDMS is stable to keep high hydrophilicity after a month of the modification. On the other hand, one of sulfobetaine polymers showed distinguished adsorption behavior in the case of polycarbonate surface treated with plasma. Suppression effect for nonspecific adsorption of BSA was evaluated using polycarbonate and acrylic resin modified with the polymers. The modified surfaces showed suppression effect for nonspecific adsorption of BSA compared with the surface only treated with plasma.

Novel pH-responsive polymeric micelles prepared through self-assembly of amphiphilic block copolymer with poly-4-vinylpyridine block synthesized by mechanochemical solid-state polymerization.

We fabricated polymeric micelles containing 5-fluorouracil (5-FU) or fluorescein using the amphiphilic block copolymer, poly-4-vinylpyridine-b-6-O-methacryloyl galactopyranose. Although the polymeric micelles were stable at pH 7.4, they readily decomposed at pH 5, resulting in near complete release of 5-FU. Uptake of polymeric micelles containing fluorescein by HepG2 and HCT116 cells was also investigated. With both cell types, strong fluorescence was observed after a 12-h incubation, but the fluorescence weakened after 24 h of incubation. The fluorescein incorporated into the polymeric micelles was released into acidic organelles (endosome and/or lysosome), from which it diffused throughout the cell. The cytotoxicity of polymeric micelles containing 5-FU was evaluated against HepG2 cells using a CCK-8 assay. The results suggest that polymeric micelles containing 5-FU are more cytotoxic to HepG2 cells than free 5-FU.

Behavior of polyethylene glycol molecules at an oscillating solid-liquid interface.

The behavior of polyethylene glycol (PEG) molecules at an oscillating solid-liquid interface was examined using a quartz crystal microbalance (QCM). The number-average molecular weights (Mn) of PEG molecules were systematically varied. This study revealed that the shift in the energy dissipation factor, ΔD, of the QCM against square root of the density-viscosity product of the PEG solution is linear and has an intercept. Moreover, the systematical analysis revealed that the ΔD slope rapidly decreases with Mn, and the ΔD intercept becomes constant above 1.1×10(4) g/mol. Those results indicated that the resonant length of PEG molecules moving with the oscillating plate at 9 MHz is 97.6 Å. We also found that the difference between the resonant lengths calculated from ΔD and series resonance-frequency shift, ΔF, is related to the density of the thin PEG layer formed on the gold electrode.

Physical properties of self-assembled monolayers of Mercapto Oligo (ethylene oxide) methyl ether on gold.

Physical properties (chemical adsorption rate, viscoelasticity, thickness, and conformation) for self-assembled monolayers (SAMs) of mercapto oligo (ethylene oxide) methyl ethers on gold were determined by quartz crystal microbalance (QCM). The values of thickness, shear viscosity and elastic shear modulus of SAMs increase with unit number of oligo (ethylene oxide) segment. However, the chemical adsorption rate determined by a Langmuir isotherm does not show such a unit number regularity. Those results suggest that difference in unit number of oligo (ethylene oxide) segment can significantly affects physical properties of SAMs.

Dynamic properties of the polyethylene glycol molecules on the oscillating solid-liquid interface.

The dynamic properties of polyethylene glycol (PEG) molecules on the solid-liquid interface oscillating at MHz were investigated using the quartz crystal microbalance (QCM). The number-average molecular weights (M(n)) of the PEG molecules were systematically varied over 4 orders of magnitude. This study makes it clear that the series-resonant frequency shift, ΔF, of the QCM against the square root of the density-viscosity product of the PEG solution is linear and has the intercept. Moreover, systematical analysis reveals that the ΔF slope rapidly decreases with M(n) and that the ΔF intercept becomes constant above 4.0×10(3) g mol(-1). As a result, those reveal that the resonant length of the PEG molecule moving with the oscillating plate of 9 MHz is 54.2 Å. We also find that the behaviors of ΔF due to M(n) are mainly caused by the length of the PEG molecule.

Noise-induced order in the chaos of the Belousov-Zhabotinsky reaction.

Noise can stabilize a metastable state in such a way that the system remains in this state for a longer time than in the absence of noise. When this phenomenon is observed in chaos, it is called "noise-induced order." We have experimentally detected noise-induced order in the Belousov-Zhabotinsky reaction. That is, when noise is added to the chaos with the flow rate near the period-three oscillation, a decrease of the maximum Lyapunov exponent and a convergence of the Fourier spectrum are observed. Moreover, the analysis on the one-dimensional return map reveals that noise-induced order is caused by the convergence of the chaotic trajectory into the laminar region.

Immersion angle dependence of the resonant-frequency shift of the quartz crystal microbalance in a liquid: effects of longitudinal wave.

We investigated the effects of the longitudinal wave on the immersion angle dependence of the resonant-frequency shift, deltaF, of the quartz crystal microbalance, QCM. In order to study exactly the effects, we employed the three types of cells: normal cell, cell with the glass beads and cell with sponge. The longitudinal wave exists in the normal cell. On the other hand, both the cell with the glass beads and the cell with sponge eliminate the longitudinal wave. As results, we have found that the tendencies of deltaF are the same in the three types of cells. That is, we conclude that the longitudinal wave does not have effects on the immersion angle dependence of deltaF.

Surface plasmon resonance-based trace detection of small molecules by competitive and signal enhancement immunoreaction.

A surface plasmon resonance (SPR)-immunosensor for detection of the low molecular weight compound 2,4-dinitorophenol (DNP) at ultra-low concentration has been developed. The sensor strategy is based on a competitive immunoreaction between DNP and a DNP-protein conjugate, namely DNP-bovine serum albumin conjugate (DNP-BSA). Anti-DNP monoclonal antibody was immobilized on a gold thin-film coated SPR-sensor chip by means of a chemical coupling process. DNP-BSA, on contact with the anti-DNP antibody immobilized SPR-immunosensor chip causes an increase in the resonance angle of the sensor chip. The optimum concentration of immobilized antibody on the SPR-sensor chip is 100 microg mL(-1). The SPR-immunosensor response for free DNP determination using the competitive immunoreaction had a response time of ca. 15 min. Using this method, DNP could be determined in the concentration range 1 ppt to 1 ppb. The SPR signal for ppt levels of DNP was enhanced by a factor of three by subsequently treating immuno-bound DNP-BSA with a secondary anti-DNP antibody.

Dependence on the electric power of the immersion-angle dependence of the resonant-frequency shift of a quartz crystal microbalance in a liquid.

We have investigated the immersion-angle dependence of the series resonant-frequency shift, DeltaF, of the quartz crystal microbalance, QCM, in a Newtonian liquid from the point of view of the supplied electric power level. In the low electric power levels, the immersion-angle dependence and the transition phenomenon of the DeltaF are observed. However, when the higher electric power levels are supplied to the QCM, the region of the transition phenomenon of the DeltaF decreases rapidly with an increase in the electric power level and disappears above 1.5 dBm. That is, above 1.5 dBm, the DeltaF values have only the values of 90 degrees immersion angle in all immersion angles. We suggest that the electric power is very important factor for the DeltaF in a liquid.

Antibody immobilization to phospholipid polymer layer on gold substrate of quartz crystal microbalance immunosensor.

To modify gold electrode for immunosensor to construct an artificial cell membrane structure, water-soluble amphiphilic phospholipid polymer, poly[2-methacryloyloxyehtyl phosphorylcholine-co-n-butyl methacrylate-co-p-nitrophenyloxycarbonyl poly(ethylene glycol) methacrylate (PMBN)] was applied. The polymer had active ester groups for immobilization of biomolecules and it was converted partially to thiol groups for binding to gold substrates. The partially thiolated PMBN was adsorbed on a gold electrode of quartz crystal microbalance (QCM). Surface characterization of adsorbed PMBN layers was thoroughly investigated with reflectance anisotropy spectroscopy, ellipsometry spectroscopy, dynamic contact angle and X-ray photoelectron spectroscopy measurements. Among several PMBN, having different degree of thiolation, it was concluded that 21.5% thiolated PMBN layer had the most well-ordered phosphorylcholine groups in its outer surface. The proteins adsorption test revealed that the phosphorylcholine group on the outer side of PMBN layers, which was substituted their active ester groups by glycine, showed suppress the non-specific adsorption of proteins, such as bovine serum albumin and gamma-globulin. Also, through antigen-antibody binding evaluation, the anti-C-reactive protein antibody immobilized on the PMBN surface worked well and it was confirmed that denaturation of the antibody on the PMBN layers was hardly occurred in spite of 60 days storage at 4 degrees C. The antibody conjugated phospholipid polymer layer with well-ordered phosphorylcholine group could be outstanding functional membrane for biomedical diagnostic devices without non-specific binding and reduction of immunologic activity of immobilized antibody.

Adsorption of 125I-labeled immunoglobulin G, its F(ab')2 and Fc fragments onto plasma-polymerized films.

Plasma-polymerized films were formed on flat glass plates using allylamine, acrylic acid, acrolein, and allylcyanide as monomers. Adsorption of (125)I-labeled-proteins such as immunoglobulin G (IgG), its F(ab')(2) and Fc fragments, and human serum albumin (HSA) was measured on these plasma-polymerized (PP) films covering the glass plates and on commercially available polymer plates. The adsorption isotherm followed the Langmuir equation, from which the binding constant and amount of saturation binding were estimated. We found that, in general, a cationic surface had higher affinity for protein adsorption than an anionic surface. Among the surfaces examined, the PP-allylamine surface showed the highest binding capacity (264.2 nmol/m(2)) for F(ab')(2) fragment: it was remarkably high. Of the surfaces examined, the PP-acrylic acid surface showed the lowest binding capacity (12.8 nmol/m(2)) for F(ab')(2) fragment. The PP-acrylic acid surface also indicated the lowest protein binding capacity for IgG (16.5 nmol/m(2)), Fc-IgG (32.4 nmol/m(2)) and HSA (16.7 nmol/m(2)), respectively. These imply that the PP-acrylic acid film is useful to fabricate as a low protein adsorption material which expected to decrease cell adhesion. Results of our investigation indicate that the plasma-polymerization technique is promising for fabrication of a smart NanoBio-interface which can control the protein adsorption on a solid-phase substrate using a suitable monomer such as allylamine for the large adsorption and acrylic acid for the small adsorption.

Characteristics of the series resonant-frequency shift of a quartz crystal microbalance in electrolyte solutions.

We report the novel characteristics of the series resonant-frequency shift, DeltaF, of a one-face sealed quartz crystal microbalance (QCM) in solutions of monovalent electrolytes and divalent electrolytes. In the present study, we used NaCl, KCl, LiCl, NaNO3 and CH3COONa as monovalent electrolytes, and MgCl2, CaCl2, MgSO4 and Na2SO4 as divalent electrolytes. These experiments reveal that, in all the solutions, the DeltaF values vary linearly with an increase in the square root of the density-viscosity product of the solutions, and the immersion angle dependence of DeltaF appears. Moreover, we have found that the intercept values of DeltaF are dependent only on the cation species, and that the slope values of DeltaF with the viscosity and density of the solutions are related only to the valence of ions. We also suggest that the DeltaF values are independent of anion species, conductivity and permittivity of the solutions.

Quartz crystal microbalance immunosensors for environmental monitoring.

This paper presents discussion of quartz crystal microbalance (QCM) immunosensors for environmental monitoring. Factors limiting the practical application of antibodies to analytical problems are also presented. Among several candidates for the QCM immunosensor device, selected QCM devices and oscillating circuits were tested thoroughly and developed to obtain highly stable and sensitive frequency signals. The biointerface of QCM immunosensor was designed and controlled to immobilize antibody on the QCM surface, to reduce non-specific binding and to suppress denaturation of immobilizing antibody by self-assembled monolayer technique and artificial phospholipid (2-methacryloyloxyethyl phosphorylcholine (MPC)) polymer. MPC polymer as a antibody-stabilizing reagent was added to reduce non-specific binding of the antigen solution and stabilize the immunologic activity of the antibody-immobilized QCM. In addition, it provides examples for detection and quantitation of environmental samples using QCM immunosensors. The analytical results for fly ash extracted samples of dioxins using the QCM immunosensor indicated a good relationship with GC/MS methods. The integrating protocols of the competitive immunoassay and signal-enhancing step are for detecting low molecular analytes with extremely low detection limits using an QCM immunosensor. Furthermore, its detect limitation was extended from 0.1 to 0.01 ng/ml by the signal-enhancing step when the anti-bisphenol-A antibody conjugated MPC polymeric nanoparticles was used. The QCM immunosensor method has demonstrated its effectiveness as an alternative screening method for environmental monitoring because these results were compared with results obtained through environmental monitoring methods such as ELISA and GC/MS.

Dioxin immunosensor using anti-2,3,7,8-TCDD antibody which was produced with mono 6-(2,3,6,7-tetrachloroxanthene-9-ylidene) hexyl succinate as a hapten.

To detect dioxin using a quartz crystal microbalance (QCM) immunosensor, anti-2,3,7,8-tetrachloro-p-dibenzodioxin (TCDD) monoclonal antibodies (MAbs) were produced as types of IgG1 and IgM, with mono 6-(2,3,6,7-tetrachloroxanthene-9-ylidene) hexyl succinate (as a hapten) conjugated with bovine serum albumin (dioxin-BSA). Furthermore, ScFv was generated from hybridoma-producing IgG1 MAb. Among these antibodies, ScFv showed excellent capability for dioxin detection using QCM immunosensors.

Piezoelectric immunosensor for bisphenol A based on signal enhancing step with 2-methacrolyloxyethyl phosphorylcholine polymeric nanoparticle.

An immunoassay in which BPA competed with a BPA-horseradish peroxidase conjugate for binding to anti-BPA antibodies, coupled to a piezoelectric (PZ) immunosensor, was able to detect 0.1 ng mL(-1) BPA. To enhance the sensitivity of the assay, we tested nanoparticles approximately 200 nm in diameter, coupled to anti-BPA antibodies, to increase the mass change on the surface of the immunosensor and thereby increase the frequency shift detected. This second step, using nanoparticles coated with anti-BPA antibodies, improved the sensitivity of the assay by approximately eight times at BPA concentrations below 10 ng mL(-1). Field emission-scanning electron microscopy (FE-SEM) showed that polymeric 2-methacrolyloxyethyl phosphorylcholine (MPC) nanoparticles coupled to antibodies remained monodisperse on the surface of the immunosensor and therefore produced stable signals in the immunosensors. Since the frequency shift detected in the assay mainly originated from the mass change on the surface of the PZ crystal, the colloidal stability of the antibody-conjugated particles used in the enhancement step played an extremely important role in achieving a stable and highly sensitive signal.

Quartz crystal microbalance immunosensor for highly sensitive 2,3,7,8-tetrachlorodibenzo-p-dioxin detection in fly ash from municipal solid waste incinerators.

A quartz crystal microbalance (QCM) immunosensor was developed for the detection of 2,3,7,8-tetrachlorodibenzo-p-dioxins (TCDD) in environmental pollutants. An anti-TCDD antibody was immobilized on the gold surface of the QCM via chemical coupling, and its immunologic activity was then maintained by treatment with an artificial stabilizing reagent such as poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate). A competitive immunoreaction with TCDD conjugated ovalbumin (TCDD-ovalbumin) was used to detect TCDD. A calibration curve was obtained through the competitive immunoreaction, and linearity was shown from 100 ng mL(-1) to 0.1 ng mL(-1). Also, the cross-reactivities of the anti-TCDD monoclonal antibody were thoroughly evaluated with several TCDD derivatives. The relationships between GC-MS, ELISA, and QCM were compared using fly ash samples from a municipal solid waste, which were prepared using an accelerated solvent extractor. For 23 samples, the experimental relationship between the TCDD concentration by QCM vs. the TCDD concentration by ELISA was y= 1.07x + 2.70, r= 0.99, and the TCDD concentration by QCM vs. the toxic equivalent quantity (TEQ) value by GC-MS was y= 2.46x - 14.98, r= 0.89.

Dynamical behavior of lipid bilayer membranes for taste substances under random membrane-potential fluctuations.

The dynamical behavior of the lipid bilayer membranes was experimentally studied under superposition of random or periodic membrane-potential fluctuations. The analysis of the mutual information has revealed that, in less than 10 Hz of random fluctuations, each of the time series of the mutual information of the transmembrane current for the five chemical substances (taste substances) has its inherent pattern, but not in a periodic fluctuation. On the other hand, the analysis of the power spectrum of the frequency could not distinguish those five basic taste substances in both random and periodic fluctuations. We provide the new detection idea of chemical substances by random fluctuations.

Evaluation of a high-affinity QCM immunosensor using antibody fragmentation and 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer.

This study evaluated construction of a highly affinitive quartz crystal microbalance (QCM) immunosensor using anti-C-reactive protein (CRP) antibody and its fragments for CRP detection. Three types of antibody were immobilized on the surface of a QCM via covalent-bounding. Then affinity was evaluated through antigen-antibody binding between CRP and its antibody. Affinity between antigen-antibody was shown to be highest when anti-CRP F(ab')2-IgG antibody (70 microg/mL) was immobilized on the QCM. In case of anti-CRP F(ab')2-IgG antibody, affinity which was attributable to antigen-antibody binding was almost twice that of anti-CRP IgG antibody, which is used conventionally for QCM immunosensors. In addition, when it was treated with 2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate, so-called MPC polymer, highly affinitive and selective immunosensing for CRP was achieved without non-specific binding from plasma proteins in human serum. When anti-CRP F(ab')2-IgG antibody was immobilized on the QCM, the detection limit and the linearity of CRP calibration curve were achieved at concentrations from 0.001 to 100 microg/dL even during investigation in serum samples. Experimental results verified the successful construction of a highly affinitive and selective QCM-immunosensor which was modified with anti-CRP F(ab')2-IgG antibody and MPC polymer.