We have established the nanofabrication technique for constructing nanopillars with high aspect ratio (100-500 nm diameter and 500-5000 nm tall) inside a microchannel on a quartz chip. The size of pillars and the spacing between pillars are designed as a DNA sieving matrix for optimal analysis of large DNA fragments over a few kilobase pairs (kbp). A chip with nanopillar channel and simple cross injector was developed based on the optimal design and applied to the separation of DNA fragments (1-38 kbp) and large DNA fragments (lambda DNA, 48.5 kbp; T4 DNA, 165.6 kbp) that are difficult to separate on conventional gel electrophoresis and capillary electrophoresis without a pulsed-field technique. DNA fragments ranging from 1 to 38 kbp were separated as clear bands, and furthermore, the mixture of lambda DNA and T4 DNA was successfully separated by a 380-microm-long nanopillar channel within only 10 s even under a direct current (dc) electric field. Theoretical plate number N of the channel (380-1450 microm long) was 1000-3000 (0.7 x 10(6)-2.1 x 10(6) plates/m). A single DNA molecule observation during electrophoresis in a nanopillar channel revealed that the optimal nanopillars induced T4 DNA to form a narrow U-shaped conformation during electrophoresis whereas lambda DNA kept a rather spherical conformation. We demonstrated that, even under a dc electric field, the optimal nanopillar dimensions depend on a gyration radius of DNA molecule that made it possible to separate large DNA fragments in a short time.

A fully packed capillary electrochromatographic (CEC) microchip showing improved solution and chip handling was developed. Microchannels for the CEC microchip were patterned on a cyclic olefin copolymer substrate by injection molding and packed fully with 0.8-microm monodisperse colloidal silica beads utilizing a self-assembly packing technique. The silica packed chip substrate was covered and thermally press-bonded. After fabrication, the chip was filled with buffer solution by self-priming capillary action. The self-assembly packing at each channel served as a built-in nanofilter allowing quick loading of samples and running buffer solution without filtration. Because of a large surface area-to-volume ratio of the silica packing, reproducible control of electroosmotic flow was possible without leveling of the solutions in the reservoirs resulting 1.3% rsd in migration rate. The capillary electrophoretic separation characteristics of the chip were studied using fluorescein isothiocyanate (FITC)-derivatized amino acids as probe molecules. A mixture of FITC and four FITC-derivatized amino acids was successfully separated with 2-mm separation channel length.

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The distribution and the orgins of substance P (SP)-positive fibers in the papillae of the rat tongue were investigated by means of the indirect immunofluorescent method.Three types of papilla contain SP-positive fibers, though the number of these fibers varies from papilla to papilla. The circumvallate papilla contains the greatest number of SP-positive fibers, followed by the foliate papillae and the fungiform papillae; the filiform papillae lack SP-positive fibers. The papillar SP-positive fibers form dense bands in the lamina propria just beneath the epithelium. Some of the fibers enter the epithelium and the taste buds. It should be stressed that not every taste bud is provided with SP-positive fibers: Only 95% of the taste buds in the foliate papillae, 70% of the taste buds in the fungiform papillae, and 40% of the taste buds in the circumvallate papillae contain detectable SP-positive fibers. Unilateral section of the glossopharyngeal nerve resulted in a complete disappearance of SP-positive fibers in the foliate papillae on the operated side, and a slight decrease in the circumvallate papillae on both sides. Bilateral section of the glossopharyngeal nerve resulted in a complete disappearance of SP-positive fibers in the foliate and circumvallate papillae. Following unilateral section of the chorda tympani, SP-positive fibers in the taste buds of the fungiform papillae disappeared completely. In addition, unilateral neurotomy of the mandibular nerve resulted in a complete disappearance of SP-positive fibers in the epithelium of the fungiform papillae. These facts strongly indicate that SP in the foliate and circumvallate papillae is supplied by the glossopharyngeal nerve, SP in the taste buds of the fungiform papillae by the chorda tympani, and SP in the epithelium of the fungiform papillae by the third division of the trigeminal nerve.Since the recognition of substance P (SP) by von Euler and Gaddum ('31), this substance has been known to be a polypeptide which might act as a neurotransmitter (see Leeman and Mroz, '74, for review). Leeman and her coAddress reprint requests to Dr.

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We have developed quartz microchips for electrophoresis and a linear imaging UV detector along with the microchip. The microchips have an optical slit, which cut off the stray light in order to improve the sensitivity of UV absorption detection on the chip, at the bonding interface. They have been successfully fabricated on synthesized quartz glass substrates using the hydrofluoric acid (HF) solution bonding method. The signal level of UV absorption detection was effectively improved by applying microchips with the "on-chip" optical slit. It is also possible to improve the signal-to-noise ratio by repetitive scanning of linear photodiode array located along the separation channel, and signal averaging during elimination of the potential. Furthermore, the analysis may be performed until the separation of the target component is complete, because the real-time migration pattern of each component in the sample can be seen just as in a slab-gel electrophoresis, thus enabling a shorter analysis time.

We have developed quartz microchips for electrophoresis and a linear imaging UV detector along with the microchip. The microchips have an optical slit, which cut off the stray light in order to improve the sensitivity of UV absorption detection on the chip, at the bonding interface. They have been successfully fabricated on synthesized quartz glass substrates using the hydrofluoric acid (HF) solution bonding method. The signal level of UV absorption detection was effectively improved by applying microchips with the "on-chip" optical slit. It is also possible to improve the signal-to-noise ratio by repetitive scanning of linear photodiode array located along the separation channel, and signal averaging during elimination of the potential. Furthermore, the analysis may be performed until the separation of the target component is complete, because the real-time migration pattern of each component in the sample can be seen just as in a slab-gel electrophoresis, thus enabling a shorter analysis time.