A microsensor for adenosine-5′-triphosphate pH-sensitive field effect transistors

The sensor for adenosine-5′-triphosphate (ATP) is based on H⁺-ATPase immobilized via a polyvinylbutyral resin on a pH-sensitive field effect transistor. A linear relationship was obtained between the initial rate of change of the differential gate output voltage and the logarithm of the ATP concentration over the range 0.2–1.0 mM ATP. The optimum pH was 9.0 at 40°C but pH 7.0 was preferred for routine measurements. Only slight responses were obtained for 1 mM glucose, creatinine or urea. The ATP-sensing system exhibited a response to 1 mM ATP for at least 18 days.     

Scanning near-field optical/atomic force microscopy for fluorescence imaging and spectroscopy of biomaterials in air and liquid: Observation of recombinantEscherichia coli with gene coding to green fluorescent protein

We have developed a system of scanning near-field optical/atomic force microscopy (SNOM/AFM) for fluorescence imaging and spectroscopy of biomaterials in air and liquid. SNOM/AFM uses a bent optical fiber simultaneously as a dynamic force AFM cantilever and a SNOM probe. Optical resolution of SNOM images shows about 50 nm in an illumination mode for a standard sample of a patterned chromium layer of 20 nm thickness on a quartz glass plate. The SNOM/AFM system contains a photon counting system and polychrometer/ICCD (intensified charge coupled device) system for observation of the fluorescence image and spectrograph of micro areas, respectively. The gene coding to green fluorescence protein (GFP) was cloned in recombinantEscherichia coli (E. coli). Topography, fluorescence image and spectrograph of recombinantE. coli by SNOM/AFM showed a difference in fluorescence in individualE. coli. Fluorescence activity of GFP can thus be used as a convenient indicator of transformation. SNOM/AFM is also applicable to observe immobilizedE. coli on a glass plate in water with a liquid chamber and may allow the viewing of observation of floating organisms.     

Formation of small single-layer and nested BN cages under electron irradiation of nanotubes and bulk material

12 N₁₂, B₁₆N₁₆ and B₂₈N₂₈ octahedra which were predicted to be magic clusters for the BN system from electronic structure calculations. Received: 2 March 1998     

Circumventing air bubbles in microfluidic systems and quantitative continuous-flow PCR applications

Polymerase chain reaction (PCR) is an essential part of research based on genomics or cell analysis. The development of a microfluidic device that would be suitable for high-temperature-based reactions therefore becomes an important contribution towards the integration of micro-total analysis systems (μTAS). However, problems associated with the generation of air bubbles in the microchannels before the introduction of the assay liquid, which we call the “initial start-up” in this study, made the flow irregular and unstable. In this report, we have tried to address these problems by adapting a novel liquid-flow method for high-temperature-based reactions. A PDMS-based microfluidic device was fabricated by soft-lithography techniques and placed on a cartridge heater. The generation of the air bubbles was prevented by introducing the fluorinated oil, an inert and highly viscous liquid, as the cap just before the introduction of the sample solutions into the microchannels. The technique was applied for continuous-flow PCR, which could perform PCR on-chip in a microfluidic system. For the evaluation of practical accuracy, plasmid DNA that serves as a reference molecule for the quantification of genetically modified (GM) maize was used as the template DNA for continuous-flow PCR. After PCR, the products were collected in a vial and analyzed by gel electrophoresis to confirm the accuracy of the results. Additionally, quantitative continuous-flow PCR was performed using TaqMan technology on our PCR device. A laser detection system was also used for the quantitative PCR method. We observed a linear relationship between the threshold cycle (Ct) and the initial DNA concentration. These results showed that it would be possible to quantify the initial copies of the template DNA on our microfluidic device. Accurate quantitative DNA analysis in microfluidic systems is required for the integration of PCR with μTAS, thus we anticipate that our device would have promising potential for applications in a wide range of research.     

Detection of Alzheimer's tau protein using localised surface plasmon resonance-based immunochip

In this study, we present the detection of tau protein, at room temperature, using a multi-spot localised surface plasmon resonance (LSPR)-based immunochip. To the best of our knowledge, this is the first report of an immunochip for tau protein. The detection method includes fabrication of a gold-capped nanoparticle LSPR chip, formation and functionalisation of a self-assembled monolayer (SAM), immobilisation of a suitable linker, effective blocking of non-specific adsorption, immobilisation of a monoclonal anti-tau antibody (tau-mAb), and finally, the optimum conditions for the immuno-reaction between tau-mAb and the antigen were determined. The method has a high performance, enables detection of tau at 10pg/mL, lower than the cut-off value of 195pg/mL (for AD) for tau protein in cerebral spinal fluid (CSF). Further, we demonstrated selectivity of the technique by showing that the introduction of bovine serum albumin (BSA), perhaps the most abundant protein component in serum and CSF, does not interfere with the detection of tau. This method also offers a potential platform for studying tau interactions with other proteins and/or potential drug candidates and could also be easily adapted for detecting phosphorylated tau and other AD biomarkers.     

Regioselective double hydrophosphination of terminal arylacetylenes catalyzed by an iron complex

The first catalytic double hydrophosphination of alkynes was achieved by reaction with diarylphosphines in the presence of an iron catalyst. The double hydrophosphination proceeded regioselectively and effectively for various secondary arylphosphines and terminal alkynes to give 1,2-bisphosphinoethane derivatives.     

A microsensor for urea based on an ion-selective field effect transistor

The pH response of a pH-sensitive field-effect transistor (FET) is not affected by a ca. 1-μm thick membrane formed from λ-aminopropyltriethoxysilane and glutaraldehyde over the gate insular by a vapor deposition method. The response between pH 5.5 and 8.5 is ca. 61 mV pH^?1 at 37°C in 5 mM Tris-HCl buffer. When urease is immobilized on the membrane, the sensor gives a linear relationship between the initial rate of the output gate voltage change and the logarithmic value of urea concentration between 16.7 and 167 mM. Determination of urea is possible within 30 s. The optimum pH is 6.0–6.5, at 37°C. The system can be used for 20 days with only slight loss of enzymatic activity.     

Piezoelectric immuno sensor for the detection of candida albicans microbes

The system consists of an AT-cut quartz piezoelectric crystal, oscillator and frequency counter. The surface of the palladium-plated electrodes is oxidized anodically, and anti-Candida antibody is immobilized onto the surface. The crystal sensor is dipped into Candida suspension and the surface mass increase, caused by immuno-adsorption of Candida, is measured by the decrease in the resonant frequency of the crystal. The frequency shift is correlated with C. albicans concentrations in the range 10⁶?5 × 10⁸ cells cm^?3. The crystal sensor showed no response to Saccharomyces cerevisiae.     

Microfabrication of encoded microparticle array for multiplexed DNA hybridization detection

A strategy for the high-sensitivity, high-selectivity, and multiplexed detection of oligonucleotide hybridizations has been developed with an encoded Ni microparticle random array that was manufactured by a "top-down" approach using micromachining and microfabrication techniques.