Integration of nanoporous membranes into microfluidic devices: electrokinetic bio-sample pre-concentration

The integration of nanoporous membranes into microfluidic devices allows a wide range of analytical and biochemical applications such as stable concentration gradient generation, sample pre-concentration, and ion and biomolecule filtration in a controllable manner. However, further applications of nanoporous membranes in microfluidic devices require rapid and controllable fabrication methods of various nanoporous precursor materials; currently, few such methods exist. Here, we describe simple and robust methods that can be used for microfabricating four different precursor materials as leakage-tight membranes in a microfluidic channel network. The methods consist of a common integration process and individual solidification processes such as solvent evaporation, UV-curing, and temperature treatment. We demonstrate that the fabricated membranes can be used for electrokinetic, nanofluidic pre-concentration of bio-samples such as proteins, cells, and microspheres on either the anodic or cathodic side of the membranes. In addition, we not only characterize the physicochemical properties of the membranes such as conductance of membrane-integrated microchannels, relative permselectivity, and pre-concentration ability, but also compare fabrication availability, membrane robustness, surface charge density tunability and biocompatibility with buffer solutions. The methods are versatile for many nanoporous precursor materials and easy to control the location and dimension of the membranes. Hence, the methods developed and the characterized properties of the membranes tested in this work could be widely employed for further applications of nanoporous membranes in microfluidic systems.close6

Cracking-assisted photolithography for mixed-scale patterning and nanofluidic applications

Cracks are observed in many environments, including walls, dried wood and even the Earth's crust, and are often thought of as an unavoidable, unwanted phenomenon. Recent research advances have demonstrated the the ability to use cracks to produce various micro and nanoscale patterns. However, patterns are usually limited by the chosen substrate material and the applied tensile stresses. Here we describe an innovative cracking-assisted nano-fabrication technique that relies only on a standard photolithography process. This novel technique produces well-controlled nanopatterns in any desired shape and in a variety of geometric dimensions, over large areas and with a high throughput. In addition, we show that mixed-scale patterns fabricated using the 'crack-photolithography' technique can be used as master moulds for replicating numerous nanofluidic devices via soft lithography, which to the best of our knowledge is a technique that has not been reported in previous studies on materials' mechanical failure, including cracking.open2

Ion concentration polarization in a single and open microchannel induced by a surface-patterned perm-selective film

We describe a novel and simple mechanism for inducing ion concentration polarization (ICP) using a surface-patterned perm-selective nanoporous film like Nafion in single, open microchannels. Such a surface-patterned Nafion film can rapidly transport only cations from the anodic side to the cathodic side through the nanopore clusters so that it is possible to generate an ICP phenomenon near the Nafion film. In this work, we characterize transport phenomena and distributions of ion concentration under various electric fields near the Nafion film and show that single-channel based ICP (SC-ICP) is affected by Nafion film thicknesses, strengths of applied electric fields, and ionic strengths of buffer solutions. We also emphasize that SC-ICP devices have several advantages over previous dual-channel ICP (DC-ICP) devices: easy and simple fabrication processes, inherently leak-tight, simple experimental setup requiring only one pair of electrodes, stable and robust ICP induced rapidly, and low electrical resistances helping to avoid Joule heating, and membrane perm-selectivity breakdown but allowing as high bulk flow as an open, plain microchannel. As an example of applications, we demonstrate that SC-ICP devices not only have high potential in pre-concentrating proteins in massively parallel microchannels but also enable the concentration and lysis of bacterial cells simultaneously and continuously on a chip; therefore, proteins within the cells are extracted, separated from the concentrated cells and then pre-concentrated at a different location that is closer to the Nafion film. Hence, we believe that the SC-ICP devices have higher possibilities of being easily integrated with traditional microfluidic systems for analytical and biotechnological applications.close10

Whole-genome association studies of alcoholism with loci linked to schizophrenia susceptibility

BACKGROUND: Alcoholism is a complex disease. There have been many reports on significant comorbidity between alcoholism and schizophrenia. For the genetic study of complex diseases, association analysis has been recommended because of its higher power than that of the linkage analysis for detecting genes with modest effects on disease. RESULTS: To identify alcoholism susceptibility loci, we performed genome-wide single-nucleotide polymorphisms (SNP) association tests, which yielded 489 significant SNPs at the 1% significance level. The association tests showed that tsc0593964 (P-value 0.000013) on chromosome 7 was most significantly associated with alcoholism. From 489 SNPs, 74 genes were identified. Among these genes, GABRA1 is a member of the same gene family with GABRA2 that was recently reported as alcoholism susceptibility gene. CONCLUSION: By comparing 74 genes to the published results of various linkage studies of schizophrenia, we identified 13 alcoholism associated genes that were located in the regions reported to be linked to schizophrenia. These 13 identified genes can be important candidate genes to study the genetic mechanism of co-occurrence of both diseases

First Best Bayesian Privatization Mechanisms

A planner is interested in designing an ex-post efficient, individually rational, Bayesian mechanism for allocating a single indivisible object to one of the agents who knows his own valuation and only the distribution of other agents' valuations of the object. In this paper, we show that it is impossible to design such a mechanism without any transfers among agents and the planner. However, we discover and describe an ex-post efficient, ex-post individually rational, Bayesian mechanism which balances transfers among agents without any payment to (or from) the planner. Our result that an ex-post efficient, ex-post individually rational, transfer balanced, Bayesian mechanism exists, is in stark contrast to two well-known impossibility results in the literature; the nonexistence of a Bayesian public good mechanism satisfying expost efficiency, individual rationality and budget balance (Laffont and Maskin (1979)) and the impossibility of an ex-post efficient, individually rational, Bayesian bilateral trading mechanism between a seller and a buyer without an outside subsidy (Myerson and Satterthwaite (1983))

Microfluidic device for analyzing preferential chemotaxis and chemoreceptor sensitivity of bacterial cells toward carbon sources

We present a novel microfluidic device that enables high sensitive analyses of the chemotactic response of motile bacterial cells (Escherichia coli) that swim toward a preferred nutrient by sorting and concentrating them. The device consists of the Y-shaped microchannel that has been widely used in chemotaxis studies to attract cells toward a high concentration and a concentrator array integrated with arrowhead-shaped ratchet structures beside the main microchannel to trap and accumulate them. Since the number of accumulated cells in the concentrator array continuously increases with time, the device makes it possible to increase the sensitivity of detecting chemotactic responses of the cells about 10 times greater than Y-shaped channel devices in 60 min. In addition, the device can characterize the relative chemotactic sensitivity of chemoreceptors to chemoeffectors by comparing the number of cells in the concentrator array at different distances from the channel junction. Since the device allows the analysis of both the chemotactic responses and the sensitivity of chemoreceptors with high resolution, we believe that not only can the device be broadly used for various microbial chemotaxis assays but it also can further the advancement of microbiology and even synthetic biology.close9

Controlled open-cell two-dimensional liquid foam generation for micro- and nanoscale patterning of materials

Liquid foam consists of liquid film networks. The films can be thinned to the nanoscale via evaporation and have potential in bottom-up material structuring applications. However, their use has been limited due to their dynamic fluidity, complex topological changes, and physical characteristics of the closed system. Here, we present a simple and versatile microfluidic approach for controlling two-dimensional liquid foam, designing not only evaporative microholes for directed drainage to generate desired film networks without topological changes for the first time, but also microposts to pin the generated films at set positions. Patterning materials in liquid is achievable using the thin films as nanoscale molds, which has additional potential through repeatable patterning on a substrate and combination with a lithographic technique. By enabling direct-writable multi-integrated patterning of various heterogeneous materials in two-dimensional or three-dimensional networked nanostructures, this technique provides novel means of nanofabrication superior to both lithographic and bottom-up state-of-the-art techniques

Non-Einstein Viscosity Phenomenon of Acrylonitrile–Butadiene–Styrene Composites Containing Lignin–Polycaprolactone Particulates Highly Dispersed by High-Shear Stress

Lignin powder was modified via ring-opening polymerization of caprolactone to form a lignin–polycaprolactone (LPCL) particulate. The LPCL particulates were mixed with an acrylonitrile–butadiene–styrene (ABS) matrix at an extremely high rotational speed of up to 3000 rpm, which was achieved by a closed-loop screw mixer and in-line melt extruder. Using this high-shear extruding mixer, the LPCL particulate size was controlled in the range of 3395 nm (conventional twin-screw extrusion) down to 638 nm (high-shear mixer of 3000 rpm) by altering the mixing speed and time. The resulting LPCL/ABS composites clearly showed non-Einstein viscosity phenomena, exhibiting reduced viscosity (2130 Pa·s) compared to the general extruded composite one (4270 Pa·s) at 1 s–1 and 210 °C. This is due to the conformational rearrangement and the increased free volume of ABS molecular chains in the vicinity of LPCL particulates. This was supported by the decreased glass transition temperature (Tg, 83.7 °C) of the LPCL/ABS composite specimens, for example, giving a 21.8% decrement compared to that (107 °C) of the neat ABS by the incorporation of 10 wt % LPCL particulates in ABS. The LPCL particulate morphology, damping characteristics, and light transmittance of the developed composites were thoroughly investigated at various levels of applied shear rates and mixing conditions. The non-Einstein rheological phenomena stemming from the incorporation of LPCL particulates suggest an interesting plasticization methodology: to improve the processability of high-loading filler/polymer composites and ultra-high molecular weight polymers that are difficult to process because of their high viscosity

A microfluidic concentrator array for quantitative predation assays of predatory microbes

We present a microfabricated concentrator array device that makes it possible to quantify the predation rate of Bdellovibrio bacteriovorus, a predatory microbe, toward its prey, Escherichia coli str. MG1655. The device can accumulate both prey and predator microbes sequentially within a series of concentrator arrays using the motility of the microbes and microfabricated arrowhead-shaped ratchet structures. Since the device can constrain both prey and predator cells within 200 pL chambers at a desired range of cell densities, it was demonstrated that the device cannot only enhance the possibility of studying predation processes/cycles directly at a single cell level but can also quantify the predation rates indirectly by measuring the time-dependent fluorescent intensity signals from the prey. Furthermore, the device can produce a wide range of initial prey to predator density ratios within various concentrator arrays through the use of microfluidic mixer structures on a single array chip, which allows us to study many different conditions with a single set of cultures, and quantitatively characterize the predation behaviour/rate. Lastly, we note that this novel concentrator array device can be a very powerful tool facilitating studies of microbial predations and microbe-microbe interaction and may be broadly used in other microbial biotechnological applications.close6