Development of a package-free transparent disposable biosensor chip for simultaneous measurements of blood constituents and investigation of its storage stability.

A package-free transparent disposable biosensor chip was developed by a screen-printing technique. The biosensor chip was fabricated by stacking a substrate with two carbon electrodes on its surface, a spacer consisting of a resist layer and an adhesive layer, and a cover. The structure of the chip keeps the interior of the reaction-detecting section airtight until use. The chip is equipped with double electrochemical measuring elements for the simultaneous measurement of multiple items, and the reagent layer was developed in sample-feeding path. The sample-inlet port and air-discharge port are simultaneously opened by longitudinally folding in two biosensor units with a notch as a boundary. Then the shape of the chip is changed to a V-shape. The reaction-detecting section of the chip has a 1.0 microl sample volume for one biosensor unit. Excellent results were obtained with the chip in initial simultaneous chronoamperometric measurements of both glucose (r=1.00) and lactate (r=0.998) in the same samples. The stability of the enzyme sensor signals of the chip was estimated at ambient atmosphere on 8 testing days during a 6-month period. The results were compared with those obtained for an unpackaged chip used as a control. The package-free chip proved to be twice as good as the control chip in terms of the reproducibility of slopes from 16 calibration curves (one calibration curve: 0, 100, 300, 500 mg dl(-1) glucose; n=3) and 4.6 times better in terms of the reproducibility of correlation coefficients from the 16 calibration curves.

Development of a self-sterilizing lancet coated with a titanium dioxide photocatalytic nano-layer for self-monitoring of blood glucose.

A photocatalyst was applied to a lancet for pricking the finger to obtain an antibacterial property. A photocatalytic and uniform nano-layer of titanium dioxide (TiO2) on the surface of the lancet (0.36 mm x 24.5 mm) was formed by sputtering and annealed for crystallization of the TiO2 layer. By elementary analysis of the TiO2 layer, titanium and oxygen were detected. Next, for the estimation of the antibacterial properties resulting from the photocatalytic effect, the lancet was packed into a capillary tube filled with a suspension of Escherichia coli K-12 (non-spore-forming bacterium), and was continuously rolled in a continuous UV-irradiation system under black-light irradiation. Distinct antibacterial effects after irradiation at 0.5 mW cm(-2) for 45 min were observed in the crystallized TiO2 layer on the lancet. Finally, lancing resistances obtained by pricking an artificial skin sheet were examined using control lancets, and lancets with an unannealed TiO2 layer or an annealed TiO2 layer. The results showed almost the same lancing resistances for the control (0.53+/-0 N, n=3) and the lancet with an annealed TiO2 layer (0.51+/-0.018 N), while the lancet with an unannealed TiO2 layer showed a high lancing resistance compared with the other lancets (0.62+/-0.05 N). In conclusion, the lancet coated with a crystallized, velvety nano-layer of TiO2 obtained by annealing had antibacterial properties and a similar lancing resistance compared with the bare lancet, and showed potential for application in monitoring blood glucose in diabetes.

High-performance genetic analysis on microfabricated capillary array electrophoresis plastic chips fabricated by injection molding.

We have developed a novel technique for mass production of microfabricated capillary array electrophoresis (mu-CAE) plastic chips for high-speed, high-throughput genetic analysis. The mu-CAE chips, containing 10 individual separation channels of 50-microm width, 50-microm depth, and a 100-microm lane-to-lane spacing at the detection region and a sacrificial channel network, were fabricated on a poly(methyl methacrylate) substrate by injection molding and then bonded manually using a pressure-sensitive sealing tape within several seconds at room temperature. The conditions for injection molding and bonding were carefully characterized to yield mu-CAE chips with well-defined channel and injection structures. A CCD camera equipped with an image intensifier was used to monitor simultaneously the separation in a 10-channel array with laser-induced fluorescence detection. High-performance electrophoretic separations of phiX174 HaeIII DNA restriction fragments and PCR products related to the human beta-globin gene and SP-B gene (the surfactant protein B) have been demonstrated on mu-CAE plastic chips using a methylcellulose sieving matrix in individual channels. The current work demonstrated greatly simplified the fabrication process as well as a detection scheme for mu-CAE chips and will bring the low-cost mass production and application of mu-CAE plastic chips for genetic analysis.