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A stable liquid/liquid optical waveguide (LLW) was formed using a sheath flow, where a 15% sodium chloride (NaCl) solution functioned as the core solution and water functioned as the cladding solution (15% NaCl/water LLW). The LLW was at least 200 mm in length. The concentration distributions of the liquid core and liquid cladding solutions in the LLW system were predicted by computational fluid dynamics (CFD) to validate the characteristics of the waveguide. The broadening of the region of the fluorescence of Rhodamine B excited by the guided light and the increase in the critical angle of the guided light with the increase in the contact time of the core and the cladding solutions were well explained by CFD calculations. However, no substantial leakage of the guided light was observed despite the considerably large change in the refractive index profile of the LLW; thus, a narrower and longer waveguide was realized.

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We investigated the shape of the liquid-liquid interface in micro counter-current flows formed within microchannels. The pressure balance at the interface was calculated based on the interface geometry. Although the shape should be an arc under laminar flow, a large deformation near the center of the microchannel was observed. In the center of the microchannel, Laplace pressure (171 - 450 Pa) was induced toward the aqueous phase. In contrast, near both sidewalls, Laplace pressure (81 - 166 Pa) was induced toward the organic phase. This result suggests that opposing flow occurs in the adjacent phases near the interface, with spiral-like flow generation.

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Understanding fluid and interfacial properties in extended nanospace (10-1000 nm) is important for recent advances of nanofluidics. We studied properties of water confined in fused-silica nanochannels of 50-1500 nm sizes with two types of cross-section: (1) square channel of nanoscale width and depth, and (2) plate channel of microscale width and nanoscale depth. Viscosity and wetting property were simultaneously measured from capillary filling controlled by megapascal external pressure. The viscosity increased in extended nanospace, while the wetting property was almost constant. Especially, water in the square nanochannels had much higher viscosity than the plate channel, which can be explained considering loosely coupled water molecules by hydrogen bond on the surface within 24 nm. This study suggests specificity of fluids two-dimensionally confined in extended nanoscale, in which the liquid is highly viscous by the specific water phase, while the wetting dynamics is governed by the well-known adsorbed water layer of several-molecules thickness.

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Highly efficient cell-free plasma separation from 200 µL of human whole blood was realized via axial migration of blood cells and cross-flow filtration in a microchip. Although various analyses of small volumes of blood have been reported, a large volume of blood is necessary for obtaining blood cells and plasma for the conventional plasma separation technique of centrifugation. A highly efficient plasma separation method using small volumes of blood without hemolysis is an important issue. We developed a plasma separation method based on a microchip with a filter, which utilizes the axial migration of blood cells observed in blood vessels. Clogging and hemolysis on the filter can be prevented by the axial migration of the blood cells. Using this method, 65% of the plasma from 200 µL of whole blood was successfully separated without hemolysis. When the plasma separation microchip interfaced with a micro-ELISA system was applied to C-reactive protein (CRP) analysis, the CRP concentration obtained by the microchip showed good correlation with that obtained by conventional centrifugation. Total analysis time, including plasma separation, was achieved in only 25 min.

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Ion behavior confined in extended nanospace (10(1)-10(3) nm) is important for nanofluidics and nanochemistry with dominant surface effects. In this paper, we developed a new measurement technique of ion distribution in the nanochannel by super-resolution-laser-induced fluorescence. Stimulated emission depletion microscopy was used to achieve a spatial resolution of 87 nm higher than the diffraction limit. Fluorescein was used for ratiometric measurement of pH with two excitation wavelengths. The pH profile in a 2D nanochannel of 410 nm width and 405 nm depth was successfully measured at an uncertainty of 0.05. The excess protons, showing lower pH than the bulk, nonuniformly distributed in the nanochannel to cancel the negative charge of glass wall, especially when the electric double layer is thick compared to the channel size. The present study first revealed the ion distribution near the surface or in the nanochannel, which is directly related to the electric double layer. In addition, the obtained proton distribution is important to understand the nanoscale water structure between single molecules and continuum phase. This technique will greatly contribute to understanding the basic science in nanoscale and interfacial dynamics, which are strongly required to develop novel miniaturized systems for biochemical analysis and further applications.

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Various separation processes have been integrated in microfluidics, such as capillary electrophoresis and chromatography, on a microchip. However, it is extremely difficult to separate a complicated biological system by conventional methods. Here, we report on a feasible structure and the culture condition of human renal proximal tubule epithelial cells (RPTECs), with the aim to construct a bioartificial renal tubule on a chip. Glass microchips and a polycarbonate membrane were sealed with no leakage after a surface modification. Furthermore, matrigel was selected as an optimized extracellular matrix (ECM) for cell-proliferation on the membrane. After culturing for 5 days, RPTECs reached confluent in the chip-membrane structure, which was confirmed by nuclei staining. So far, we have constructed the basic structure and cell proliferation circumstance for the future demonstration of the RPTECs separating function. This separation microdevice has promising potential to be applied as both a unit of a circulation cell culture system and a research platform of cell biology.

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We established a confluent cardiomyocyte culture method using an 800-µm diameter cylindrical microchannel in this report. This was realized by introducing cardiomyocytes 2 times before and after turning over a microchip. The optimum condition was starting the flowing medium 2.0 h after seeding and flowing the medium at 1.0 µL/min. By applying this technology to a cardiomyocyte-based spherical heart pump device, one may develop self-fluid regulated devices that could be applied for implantable or circulation analysis device on a chip.

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We developed a novel microfluidic system, termed a micro-droplet collider, by utilizing the spatial-temporal localized liquid energy to realize chemical processes, which achieved rapid mixing between droplets having a large volume ratio by collision. In this paper, in order to clarify the characteristics of the micro-droplet collider, dynamics of droplet acceleration, stationary motion and collision in the gas phase in a microchannel were experimentally investigated with visualized images using a microscope equipped with a high-speed camera. The maximum velocity of 450 mm s(-1) and acceleration of 1500 m s(-2) of a 1.6 nL water droplet were achieved at an air pressure of 100 kPa. Measurement results of dynamic contact angles of droplets indicated that wettability of the surface played an important role in the stability of droplet acceleration and collision. We found that the bullet droplet penetrated into the target droplet at collision, which differed from bulk scale. The deformation of the droplet was strongly suppressed by the channel structure, thus stable collision and efficient utilization of the droplet energy were possible. These results are useful for estimating the localized energy, for improving the system in order to realize extreme performance, and for extending the applications of microfluidic devices.

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Recently, integrated chemical systems have been further downscaled to the 10(1)-10(3) nm scale, which we call extended-nano space. The extended-nano space is a transient space from single molecules to bulk condensed phase, and fluidics and chemistry have not been explored. One of the reasons is the lack of research tools for the extended-nano space, because the space locates the gap between the conventional nanotechnology (10(0)-10(1) nm) and microtechnology (>1 microm). For these purposes, basic methodologies were developed such as nanofabrication, fluidic control, detection methods, and surface modification methods. Especially, fluidic control is one of the important methods. By utilizing the methodologies, new specific phenomena in fluidics and chemistry were reported, and the new phenomena are increasingly applied to unique applications. Microfluidic technologies are now entering new research phase combined with the nanofluidic technologies. In this review, we mainly focus on pressure-driven or shear-driven extended-nano fluidic systems and illustrate the basic nanofluidics and the representative applications.

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Cells are frequently exploited as processing components for integrated chemical systems, such as biochemical reactors and bioassay systems. By culturing vascular endothelial cells (ECs) in integrated chemical devices, vascular models have also been fabricated. Here, we utilized a thermally fused-glass microchip which is chemically and physically stable and favorable for optical detections, and cultured human arterial ECs (HAECs) in it. HAECs reached confluence within 4 days. Survival and tolerance for high shear stress (25 dyn/cm2) of the HAECs were confirmed. Furthermore, HAECs responded to inflammatory cytokine, tumor necrosis facor-alpha (TNF-alpha) and attached to more leukocyte cell line, HL-60 cells than unstimulated HAECs. Our developed device can be applied as a human arterial model, and we propose it as a new method for vascular studies.

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Flow velocity profiles of micro counter-current flow of aqueous and butylacetate phases in a microchannel having a width of 100 microm were measured by micro particle image velocimetry. In order to analyze the hydrodynamic characteristics of the counter-current flow, we derived a simple analytical model for the velocity profile. When flow rates of the aqueous and organic phases were 0.2 and 0.1 microl/min, the model agreed well with the experimental results. Predictions about the velocity profile will contribute to estimation of the extraction efficiency in the co-current and counter-current extraction process.

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Liposomes are phospholipid vesicles that can serve as carriers of biologically active agents in vitro and in vivo. Here, we describe the movement of liposomes suspended with blood flowing in capillaries. Liposomes were coated with a polymer to extend their lifespan in rat mesenteric blood vessels and detected by fluorescent staining. Liposome activity was observed by intravital microscopy using a high-speed camera system at 5 and 60 min after liposome administration. Liposome velocity was determined using two-dimensional cross-correlation, and blood flow was measured by high-resolution PIV (particle image velocimetry). The results showed that the motion of polymer-coated liposome followed the phase averaged velocity distribution of heartbeats while flowing with red blood cells in microvessels. Liposome particles tend to move toward the near blood vessel wall in the low velocity of blood flow.

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A new technique using particle image velocimetry (PIV) has been developed to evaluate the detailed velocity profiles of red cells flowing in microvessels. The microcirculation in rat mesentery was directly observed using high-speed videomicroscopy, and the images of red cells flowing in the mesenteric arterioles were recorded simultaneously with the arterial blood pressure. Based on the high-speed videomicroscopic images obtained, velocity vectors in single or branched arterioles were evaluated to obtain velocity profiles across the cross-section of arterioles. It was shown that in single and straight arterioles the velocity profile was blunt with a pit at the central region, and its pit was marked in bifurcation. The present technique enables us to analyze red cell velocity profiles up to 0.8 microm in the spatial resolution and 1 msec in the time interval.

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Because endothelial cells are subject to flow shear stress, it is important to determine the velocity distribution in microvessels during studies of the mechanical interactions between the blood and the endothelium. Particle image velocimetry (PIV) is a quantitative method for measuring velocity fields instantaneously in experimental fluid mechanics. The authors have developed a high-resolution PIV technique that improves the dynamic flow range, spatial resolution, and measurement accuracy. The proposed method was applied to images of the arteriole in the rat mesentery, using an intravital microscope and high-speed digital video system. Taking the mesentery motion into account, the PIV technique was improved to measure red blood cell (RBC) velocity. Velocity distributions with spatial resolutions of 0.8 3 0.8 mm were obtained even near the wall in the center plane of the arteriole. The arteriole velocity profile was blunt in the center region of the vessel cross-section and sharp in the near-wall region. Typical flow features for non-Newtonian fluid are shown.

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As endothelial cells are subject to flow shear stress, it is important to determine the detailed velocity distribution in microvessels in the study of mechanical interactions between blood and endothelium. Recently, particle image velocimetry (PIV) has been proposed as a quantitative method of measuring velocity fields instantaneously in experimental fluid mechanics. The authors have developed a highly accurate PIV technique with improved dynamic range. spatial resolution and measurement accuracy. In this paper, the proposed method was applied to images of the arteriole in the rat mesentery using an intravital microscope and high-speed digital video system. Taking the mesentery motion into account, the PIV technique was improved to measure red blood cell (RBC) velocity. Velocity distributions with spatial resolutions of 0.8 x 0.8 microm were obtained even near the wall in the centre plane of the arteriole. The arteriole velocity profile was blunt in the centre region of the vessel cross-section and sharp in the near-wall region. Typical flow features for non-Newtonian fluid were shown. Time-averaged velocity profiles in six cross sections with different diameters were compared.

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