A fluorescence based method for the quantification of surface functional groups in closed micro- and nanofluidic channels

Surface analysis is critical for the validation of microfluidic surface modifications for biology, chemistry, and physics applications. However, until now quantitative analytical methods have mostly been focused on open surfaces. Here, we present a new fluorescence imaging method to directly measure the surface coverage of functional groups inside assembled microchannels over a wide dynamic range. A key advance of our work is the elimination of self-quenching to obtain a linear signal even with a high density of functional groups. This method is applied to image the density and monitor the stability of vapor deposited silane layers in bonded silicon/glass micro- and nanochannels.     

Rapid fabrication of a microdevice with concave microwells and its application in embryoid body formation

Here, we report a novel method for the fabrication of polydimethylsiloxane microdevices with complicated 3-D structures, such as concave and crater shapes, using an easily machined polymethyl methacrylate mold combined with a one-step molding process. The procedure presented here enables rapid preparation of complex 3-D microstructures varying in shape and dimensions. To regulate embryoid body (EB) formation, we fabricated a microfluidic device with an array of concave microwells and found that EBs growing in microwells maintained their shape, viability, and a high degree of homogeneity. We believe that this novel method provides an alternative for rapid prototyping, especially in fabricating devices with curved 3-D microstructures.     

Distribution and density functions of probability of error over slowly fading channels with diversity combining

In this letter, the distribution and density functions of the probability of error for Rayleigh and Rician fading channels with L-fold diversity are derived and plotted for various values of mean signal energy-to-noise ratio.     

Scalable fabrication of printed Zn//MnO2 planar micro-batteries with high volumetric energy density and exceptional safety

The rapid development of printed and microscale electronics imminently requires compatible micro-batteries (MBs) with high performance, applicable scalability, and exceptional safety, but faces great challenges from the ever-reported stacked geometry. Herein the first printed planar prototype of aqueous-based, high-safety Zn//MnO₂ MBs, with outstanding performance, aesthetic diversity, flexibility and modularization, is demonstrated, based on interdigital patterns of Zn ink as anode and MnO₂ ink as cathode, with high-conducting graphene ink as a metal-free current collector, fabricated by an industrially scalable screen-printing technique. The planar separator-free Zn//MnO₂ MBs, tested in neutral aqueous electrolyte, deliver a high volumetric capacity of 19.3 mAh/cm³ (corresponding to 393 mAh/g) at 7.5 mA/cm³, and notable volumetric energy density of 17.3 mWh/cm³, outperforming lithium thin-film batteries (≤10 mWh/cm³). Furthermore, our Zn//MnO₂ MBs present long-term cyclability having a high capacity retention of 83.9% after 1300 cycles at 5 C, which is superior to stacked Zn//MnO₂ batteries previously reported. Also, Zn//MnO₂ planar MBs exhibit exceptional flexibility without observable capacity decay under serious deformation, and remarkably serial and parallel integration of constructing bipolar cells with high voltage and capacity output. Therefore, low-cost, environmentally benign Zn//MnO₂ MBs with in-plane geometry possess huge potential as high-energy, safe, scalable and flexible microscale power sources for direction integration with printed electronics.     

益气活血、化痰通络法治疗椎-基底动脉供血不足性眩晕的临床研究

目的:观察中药益气活血、化痰通络法治疗椎-基底动脉供血不足(VBI)性眩晕的临床疗效,从血液流变学、头颅多普勒超声的相关指标改变,进一步研究其作用机理。方法:将符合纳入标准的100例患者随机分为两组。治疗组50例,口服益气活血、化痰通络中药;对照组50例,口服西药:甲磺酸倍他司汀片12mg,一天3次。两周为一疗程。分别比较两组治疗前后临床疗效、中医症候,并检测血液流变学指标及头颅多普勒超声的变化。结果:(1)临床疗效:治疗组总有效率为90.00%,明显优于对照组的76.00%(P0.05)。(2)中医症候:治疗组总有效率94%,对照组总有效率80%,治疗组对中医证候的改善明显优于对照组。(3)血液流变学:对血液流变学各指标的降低治疗组明显优于对照组(P0.05)。(4)经颅多普勒超声:治疗组对平均血流速度的提高明显优于对照组(P0.05)。结论:益气活血、化痰通络方药治疗VBI性眩晕,服用方便、安全,疗效确切,值得临床推广。     

Enhancing conjugation rate of antibodies to carboxylates: Numerical modeling of conjugation kinetics in microfluidic channels and characterization of chemical over-exposure in conventional protocols by quartz crystal microbalance

This research reports an improved conjugation process for immobilization of antibodies on carboxyl ended self-assembled monolayers (SAMs). The kinetics of antibody/SAM binding in microfluidic heterogeneous immunoassays has been studied through numerical simulation and experiments. Through numerical simulations, the mass transport of reacting species, namely, antibodies and crosslinking reagent, is related to the available surface concentration of carboxyl ended SAMs in a microchannel. In the bulk flow, the mass transport equation (diffusion and convection) is coupled to the surface reaction between the antibodies and SAM. The model developed is employed to study the effect of the flow rate, conjugating reagents concentration, and height of the microchannel. Dimensionless groups, such as the Damköhler number, are used to compare the reaction and fluidic phenomena present and justify the kinetic trends observed. Based on the model predictions, the conventional conjugation protocol is modified to increase the yield of conjugation reaction. A quartz crystal microbalance device is implemented to examine the resulting surface density of antibodies. As a result, an increase in surface density from 321 ng/cm², in the conventional protocol, to 617 ng/cm² in the modified protocol is observed, which is quite promising for (bio-) sensing applications.Microfluidics have been implemented in various bio-medical diagnostic applications, such as immunosensors and molecular diagnostic devices.¹ In the last decade, a vast number of biochemical species has been detected by microfluidic-based immunosensors. Immunosensors are sensitive transducers which translate the antibody-antigen reaction to physical signals. The detection in an immunosensor is performed through immobilization of an antibody that is specific to the analyte of interest.² The antibody is often bound to the transducing surface of the sensor covered by self-assembled monolayers (SAMs). SAMs are organic materials that form a thin, packed and robust interface on the surface of noble metals like that of gold, suitable for biosensing applications.³ Thiolic SAMs have a “head” group that shows a high affinity to being chemisorbed onto a substrate, typically gold. The SAMs'' carboxylic functional group of the “tail” end can be linked to an amine terminal of an antibody to form a SAM/antibody conjugation.^3,4 The conjugation process is usually accomplished in the presence of carbodiimides, such as 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC). A yield increasing additive, N-Hydroxysuccinimide (NHS), is often used to enhance the surface loading density of the antibody.^4,5A typical reaction for coupling the carboxylic acid groups of SAMs with the amine residue of antibodies in the presence of EDC/NHS is depicted in Figure ​Figure11.⁴ NHS promotes the generation of an active NHS ester (k₂ reaction path). The NHS ester is capable of efficient acylation of amines, including antibodies (k₃ reaction path). As a result, the amide bond formation reaction, which typically does not progress efficiently, can be enhanced using NHS as a catalyst.⁴Open in a separate windowFIG. 1.NHS catalyzed conjugation of antibodies to carboxylic-acid ended SAMs through EDC mediation (Adapted from G. T. Hermanson, Bioconjugate Techniques, 2nd. Edition. Copyright 2008 by Elsevier⁴). EDC reacts with the carboxylic acid and forms o-acylisourea, a highly reactive chemical that reacts with NHS and forms an NHS ester, which quickly reacts with an amine (i.e., antibody) to form an amide.A number of groups have studied EDC/NHS mediated conjugation reactions such as the ones depicted in Figure ​Figure1.1. The general stoichiometry of the reaction involves a carboxylic acid (SAM), an amine (antibody), and EDC to produce the final amide (antibody conjugated SAM) and urea. However, the recommended concentration ratio of the crosslinking reagents inside the buffer, i.e., the ratio of EDC and NHS with respect to adsorbates and each other, varies from one study to another.⁶ The kinetics of the reactions outlined in Figure ​Figure11 have also been investigated,^4,6–8 but only in the absence of NHS for EDC or carboxylic acids in aqueous solutions.⁸ A relatively recent experimental study verified the catalytic role of the yield-increasing reagent N-hydroxybenzotriazole (HOBt), which acts similarly to NHS.⁷ In this study, the amide formation rate (k₃ reaction path, Figure ​Figure1)1) was found to be dependent on the concentration of the carboxylic acid and EDC in the buffer solution, and independent of the amine and catalyst reagent concentration. The same group also showed that the amide bond formation kinetics is controlled by the reaction between the carboxylic acid and the EDC to give the O-acylisourea, which they marked as the rate-determining step (k₁ reaction path, Figure ​Figure11).The k₁ reaction path, or the conjugation reaction, is usually a lengthy process and takes between 1 and 3 h.^4,9 Compared to k₁, the k₂ and ?k₃ reactions are considerably faster. Microfluidics has the potential to enhance the kinetics of these reactions using the flow-through mode.^10,11 This improvement occurs because while conventional methods rely only on diffusion as the primary reagent transport mode, microfluidics adds convection to better replenish the reagents to the reaction surfaces. However, there are many fundamental fluidic and geometrical parameters that might affect the process time and reagents consumption in a microfluidics environment, such as concentration of antibodies and reagents, flow rate, channel height, and final surface density of antibodies. A model that studies the kinetics of conjugation reaction against all these parameters would therefore be helpful for the optimization of this enhanced kinetics.There are a number of reports on numerical examination of the kinetics of binding reactions in microfluidic immunoassays.^12–15 All these models developed so far couple the transport of reagents, by diffusion and convection, to the binding on the reaction surface. Myszka''s model assumes a spatially homogeneous constant concentration of reagents throughout the reaction chamber, thus fails to describe highly transport-limited conditions due to the presence of spatial heterogeneity and depletion of the bulk fluid from reagents.^16,17 In transport-limited conditions, the strength of reaction is superior to the rate of transport of reagents to the reaction surface.^18,19 More recently, the convection effects were included in a number of studies, describing the whole kinetic spectrum from reaction-limited conditions to transport-limited reactions.^20–22 Immunoreaction kinetics has also been examined with a variety of fluid driving forces, from capillary-driven flows,²⁰ to electrokinetic flows in micro-reaction patches,²¹ pressure-driven flows in a variety of geometric designs.²² Despite these comprehensive numerical investigations, the fundamental interrelations between the constitutive kinetic parameters, such as concentration, flow velocity, microchannel height, and antibody loading density, have not been studied in detail. In addition, the conjugation kinetics has not yet been exclusively examined.In this paper, a previous model for immunoreaction is modified to study the antibody/SAM conjugation reaction in a microfluidic system. Model findings are used to examine the process times recommended in the literature and possible modification scenarios are proposed. The new model connects the convective and diffusive transport of reagents in the bulk fluid to their surface reaction. The conjugation reaction is studied against fluidic and geometrical parameters such as flow rate, concentration, microchannel height and surface density of antibodies. Damköhler number is used to compare the reaction and fluidic phenomena present and justify the kinetic trends observed. Model predictions are discussed and the main finding on possible overexposure of carboxylates to crosslinking reagents, in conventional protocols, is verified by comparing the resultant antibody loading densities obtained using a quartz crystal microbalance (QCM) set up. The results demonstrate an improved receptor (antibody) loading density which is quite promising for a number of (bio-) sensing applications.^23,24 Major application areas include antibody-based sensors for on-site, rapid, and sensitive analysis of pathogens such as Bacillus anthracis,²³ Escherichia coli, and Listeria monocytogenes, and toxins such as fungal pathogens, viruses, mycotoxins, marine toxins, and parasites.²⁴     

原子荧光光谱法测定氧化铁皮中砷汞含量

用王水浸取样品中的砷、汞,用预还原剂(硫脲+抗坏血酸)还原,在用原子荧光测定溶液中砷汞元素含量,本法通过选择王水,并在水浴条件下分解样品,极大的提高了样品分解速度,缩短样品检测周期,利用原子荧光分光光度计测定,避免了冷原子吸收的不稳定性,与其他方法比较,本法操作简单,检测周期短,检测结果更加准确可靠。     

An integrated framework for RFID adoption and diffusion with a stage-scale-scope cubicle model: A case of Indonesia

This study presents a comprehensive framework to identify dynamic radio frequency identification (RFID) adoption and diffusion from three different perspectives: stages of adoption, levels of analysis, and domain of issues. The main concern of this study is stages of adoption, which covers three phases in respect of the maturity of the RFID project and the sophistication of business applications and RFID technology. The level of analysis involves different units of analysis beyond the organization level, including the industry- and country-level, which is lacking in the current literature. To understand dimensions of RFID issues, a strategy, technology, organization, people and environment (STOPE) based approach was applied. An extensive review of prior literature was conducted to find various RFID success factors and the Delphi method was applied to find positions of these factors within the framework. Based on the Delphi, some factors belong to early stage of adoption, and some others persist in the later stages of adoption. At the country level, factors such as RFID national policy, R&D policy and income per capita were accepted by most experts at the preliminary and intermediate stage; strategy and environment were accepted as important domains. To find practical implications of the framework, a case study of Indonesia was conducted at each level of analysis. Analytic hierarchy process (AHP) was applied to identify most important factors and important domains of issues with respect to the factors from the Delphi results. At country level, RFID policy, vision of leadership and RFID potential market are the most important factors; strategy and environment are the most important domains.     

Electrokinetic trapping and surface enhanced Raman scattering detection of biomolecules using optofluidic device integrated with a microneedles array

In this study, microneedles which possess sharp tips were utilized to trap and detect the biomolecules. Owing to the large curvature, the tips of the microneedles created a substantially high gradient of electric field under the non-uniform electric field which served as not only the trapping sites but also the substrate for surface enhanced Raman scattering (SERS). Separation of polystyrene microparticles with different sizes and two kinds of biomolecules (Staphylococcus aureus (S. aureus) and the red blood cells (RBCs)) were demonstrated. Moreover, in situ detection of S. aureus was performed immediately after separation was completed. The results showed that, after 15 s of sample collection, the Raman signals of S. aureus were detected and greatly enhanced through SERS effect.     

Reconfigurable microfluidic device with integrated antibody arrays for capture,multiplexed stimulation,and cytokine profiling of human monocytes

Monocytes represent a class of immune cells that play a key role in the innate and adaptive immune response against infections. One mechanism employed by monocytes for sensing foreign antigens is via toll-like receptors (TLRs)—transmembrane proteins that distinguish classes of foreign pathogens, for example, bacteria (TLR4, 5, and 9) vs. fungi (TLR2) vs. viruses (TLR3, 7, and 8). Binding of antigens activates a signaling cascade through TLR receptors that culminate in secretion of inflammatory cytokines. Detection of these cytokines can provide valuable clinical data for drug developers and disease investigations, but this usually requires a large sample volume and can be technically inefficient with traditional techniques such as flow cytometry, enzyme-linked immunosorbent assay, or luminex. This paper describes an approach whereby antibody arrays for capturing cells and secreted cytokines are encapsulated within a microfluidic device that can be reconfigured to operate in serial or parallel mode. In serial mode, the device represents one long channel that may be perfused with a small volume of minimally processed blood. Once monocytes are captured onto antibody spots imprinted into the floor of the device, the straight channel is reconfigured to form nine individually perfusable chambers. To prove this concept, the microfluidic platform was used to capture monocytes from minimally processed human blood in serial mode and then to stimulate monocytes with different TLR agonists in parallel mode. Three cytokines, tumor necrosis factor-α, interleukin (IL)-6, and IL-10, were detected using anti-cytokine antibody arrays integrated into each of the six chambers. We foresee further use of this device in applications such as pediatric immunology or drug/vaccine testing where it is important to balance small sample volume with the need for high information content.     

四川会理铅锌矿高碱度含泥尾矿氧化锌浮选生产实践

四川会理某铅锌矿选厂选别的是以铜铅锌银等硫化矿为主的多金属矿石,近年来,入选原矿含泥重、氧化率升高,选别指标有所下降,特别是选别后的硫化锌尾矿锌偏高,导致选矿的锌回收率大幅下降,因此对硫化锌尾矿氧化锌进行了再选。生产实践表明,高含泥低品位的氧化锌矿采用硫化后胺法浮选的方法时,脱泥作业至关重要。适当增加硫化时间有助于提高回收率。p H值太高不利于氧化锌的选别。     

Approaching 18% efficiency of ternary organic photovoltaics with wide bandgap polymer donor and well compatible Y6 : Y6-1O as acceptor

A series of ternary organic photovoltaics (OPVs) are fabricated with one wide bandgap polymer D18-Cl as donor, and well compatible Y6 and Y6-1O as acceptor. The open-circuit-voltage (V_OC) of ternary OPVs is monotonously increased along with the incorporation of Y6-1O, indicating that the alloy state should be formed between Y6 and Y6-1O due to their excellent compatibility. The energy loss can be minimized by incorporating Y6-1O, leading to the V_OC improvement of ternary OPVs. By finely adjusting the Y6-1O content, a power conversion efficiency of 17.91% is achieved in the optimal ternary OPVs with 30 wt% Y6-1O in acceptors, resulting from synchronously improved short-circuit-current density (J_SC) of 25.87 mA cm⁻², fill factor (FF) of 76.92% and V_OC of 0.900 V in comparison with those of D18-Cl : Y6 binary OPVs. The J_SC and FF improvement of ternary OPVs should be ascribed to comprehensively optimal photon harvesting, exciton dissociation and charge transport in ternary active layers. The more efficient charge separation and transport process in ternary active layers can be confirmed by the magneto-photocurrent and impedance spectroscopy experimental results, respectively. This work provides new insight into constructing highly efficient ternary OPVs with well compatible Y6 and its derivative as acceptor.     

Formation of lipid bilayer membrane in a poly(dimethylsiloxane) microchip integrated with a stacked polycarbonate membrane support and an on-site nanoinjector

This paper describes a new and facile approach for the formation of pore-spanning bilayer lipid membranes (BLMs) within a poly(dimethylsiloxane) (PDMS) microfluidic device. Commercially, readily available polycarbonate (PC) membranes are employed for the support of BLMs. PC sheets with 5 μm, 2 μm, and 0.4 μm pore diameters, respectively, are thermally bonded into a multilayer-stack, reducing the pore density of 0.4 μm-pore PC by a factor of 200. The BLMs on this support are considerably stable (a mean lifetime: 17 h). This multilayer-stack PC (MSPC) membrane is integrated into the PDMS chip by an epoxy bonding method developed to secure durable bonding under the use of organic solvents. The microchip has a special channel for guiding a micropipette in the proximity of the MSPC support. With this on-site injection technique, tens to hundreds of nanoliters of solutions can be directly dispensed to the support. Incorporating gramicidin ion channels into BLMs on the MSPC support has confirmed the formation of single BLMs, which is based on the observation from current signals of 20 pS conductance that is typical to single channel opening. Based on the bilayer capacitance (1.4 pF), about 15% of through pores across the MSPC membrane are estimated to be covered with BLMs.     

Co-operative R&D: why and with whom?: An integrated framework of analysis

Firms use R&D partnerships to access knowledge and build global R&D networks. This article develops an integrated framework to examine the determinants of the choice of partners with which firms co-operate on R&D. This resource-based perspective underscores the interactions between three major questions: why co-operate, who does and with whom? It argues in particular that the choice of partners is dictated by the complementary resources which the latter command. The framework is then expanded to predict the relative efficiency of R&D co-operation with different partners, including suppliers, clients, rivals, academic institutions and foreign firms. The empirical analysis, which is based on responses to France’s version of the second European community innovation survey (CIS-2), strongly supports the overall framework of analysis.     

A novel on-chip three-dimensional micromachined calorimeter with fully enclosed and suspended thin-film chamber for thermal characterization of liquid samples

A microfabricated calorimeter (μ-calorimeter) with an enclosed reaction chamber is presented. The 3D micromachined reaction chamber is capable of analyzing liquid samples with volume of 200 nl. The thin film low-stress silicon nitride membrane is used to reduce thermal mass of the calorimeter and increase the sensitivity of system. The μ-calorimeter has been designed to perform DC and AC calorimetry, thermal wave analysis, and differential scanning calorimetry. The μ-calorimeter fabricated with an integrated heater and a temperature sensor on opposite sides of the reaction chamber allows to perform thermal diffusivity and specific heat measurements on liquid samples with same device. Measurement results for diffusivity and heat capacitance using time delay method and thermal wave analysis are presented.