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Paraphrase detection is an important task in text analytics with numerous applications such as plagiarism detection, duplicate question identification, and enhanced customer support helpdesks. Deep models have been proposed for representing and classifying paraphrases. These models, however, require large quantities of human-labeled data, which is expensive to obtain. In this work, we present a data augmentation strategy and a multi-cascaded model for improved paraphrase detection in short texts. Our data augmentation strategy considers the notions of paraphrases and non-paraphrases as binary relations over the set of texts. Subsequently, it uses graph theoretic concepts to efficiently generate additional paraphrase and non-paraphrase pairs in a sound manner. Our multi-cascaded model employs three supervised feature learners (cascades) based on CNN and LSTM networks with and without soft-attention. The learned features, together with hand-crafted linguistic features, are then forwarded to a discriminator network for final classification. Our model is both wide and deep and provides greater robustness across clean and noisy short texts. We evaluate our approach on three benchmark datasets and show that it produces a comparable or state-of-the-art performance on all three.  相似文献   

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This research presents an enhanced approach for Aspect-Based Sentiment Analysis (ABSA) of Hotels’ Arabic reviews using supervised machine learning. The proposed approach employs a state-of-the-art research of training a set of classifiers with morphological, syntactic, and semantic features to address the research tasks namely: (a) T1:Aspect Category Identification, (b) T2:Opinion Target Expression (OTE) Extraction, and (c) T3: Sentiment Polarity Identification. Employed classifiers include Naïve Bayes, Bayes Networks, Decision Tree, K-Nearest Neighbor (K-NN), and Support-Vector Machine (SVM).The approach was evaluated using a reference dataset based on Semantic Evaluation 2016 workshop (SemEval-2016: Task-5). Results show that the supervised learning approach outperforms related work evaluated using the same dataset. More precisely, evaluation results show that all classifiers in the proposed approach outperform the baseline approach, and the overall enhancement for the best performing classifier (SVM) is around 53% for T1, around 59% for T2, and around 19% in T3.  相似文献   

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This work deals with the problem of optimal residual generation for fault detection (FD) in linear discrete time-varying (LDTV) systems subject to uncertain observations. By introducing a generalized fault detection filter (FDF) with four parameter matrices as the residual generator, a novel FDF design scheme is formulated as two bi-objective optimization problems such that the sensitivity of residual to fault is enhanced and the robustness of residual to unknown input is simultaneously strengthened. A generalized operator based optimization approach is proposed to deduce solutions to the corresponding optimization problems in operator forms, where the related H∞/H∞ or H?/H∞ FD performance index is maximized. With the aid of the addressed methods, the connections among the derived solutions are explicitly announced. The parameter matrices of the FDF are analytically derived via solving simple matrix equations recursively. It is revealed that our proposed results establish an operator-based framework of optimal residual generation for some kinds of linear discrete-time systems. Illustrative examples are given to show the applicability and effectiveness of the proposed methods.  相似文献   

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The disambiguation of abbreviations is a crucial step in medical knowledge organization. In the past, most scholars have focused on the problem of disambiguating medical abbreviations in single sentences; they have not systematically considered full-article abbreviation disambiguation tasks. In this work, we present a research framework for full-article medical abbreviation disambiguation (FMADRF) based on the structural characteristics of abbreviation–definition pairs in a full scientific medical article. Our method utilizes the information including context semantic information, external linguistic features, and the mapping relationships and structural similarities between abbreviations and their expansions. The model includes a four-pronged approach, identification of abbreviations and abbreviation–definition pairs, alignment and complementation of abbreviations and abbreviation expansions. The results show that our novel BBF-BLC-R model improves the recognition and modification effects of abbreviation–definition pairs, achieving the best F1 score of 91.83%. Furthermore, our new strategy combines semantic and structural information to significantly improve the effects of term alignment, with an F1 score of 97.11%. In our test, a thesaurus of abbreviations and their expansions was constructed from 13,472 full-text medical articles, resulting in 14,742 abbreviations, with 31,327 corresponding expansions. This work enhances the semantic association of terms in full medical texts, eliminating the problems of “rich” semantics and association–relation roadblocks caused by term misalignments. It further provides technical and methodological support for the organization of medical knowledge, facilitating the deep knowledge-mining capabilities of full-text medical articles.  相似文献   

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In this paper, the issues of finite-time extended dissipative analysis and non-fragile control are investigated for a class of uncertain discrete time switched linear systems. Based on average dwell-time approach, sufficient conditions for the finite-time boundedness and finite-time extended dissipative performance of the considered systems are proposed by solving some linear matrix inequalities, where using the concept of extended dissipative, we can solve the H∞, L2?L∞, Passivity and (Q, S, R)-dissipativity performance in a unified framework. Furthermore, two form of non-fragile state feedback controllers are designed to guarantee that the closed-loop systems satisfy the finite-time extended dissipative performance. Finally, simulation example is given to show the efficiency of the proposed methods.  相似文献   

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This work investigates the improved stability conditions for linear systems with time-varying delays via various augmented approaches. Some augmented approaches are augmented Lyapunov-Krasovskii functionals, augmented zero equalities, and the augmented zero equality approach. At first, by constructing augmented Lyapunov-Krasovskii functionals including the state vectors which were not considered in the previous works and augmented zero equalities, a stability criterion is proposed in the forms of linear matrix inequalities. Through the proposed Lyapunov-Krasovskii functionals and an additional functional derived from the integral inequality, a slightly improved result is derived. The proposed results do not consider the increase in the computational complexity to achieve a larger delay bound. So, by applying the augmented zero equality approach, which is a method of grafting the proposed augmented zero equality proposed in Finsler Lemma, to the proposed result, an enhanced stability result was derived. Also, the computational complexity is reduced by appropriately adjusting any vector of the integral inequality utilized in the proposed criteria. By applying some numerical examples to the proposed conditions, the effectiveness and superiority of the results are confirmed.  相似文献   

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Plagiarism remains at the top in terms of interest to the scientific community. In its many vicious forms, patchwork plagiarism is characterized by numerous unresolved issues and often passes “below the radar” of editors and reviewers. The problem of detecting the complexity of misconduct has been partially resolved by plagiarism detection software. However, interpretation of relevant reports is not always obvious or easy. This article deals with plagiarism in general and patchwork plagiarism in particular, as well as related problems that editors must deal with to maintain the integrity of scientific journals.  相似文献   

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Textual entailment is a task for which the application of supervised learning mechanisms has received considerable attention as driven by successive Recognizing Data Entailment data challenges. We developed a linguistic analysis framework in which a number of similarity/dissimilarity features are extracted for each entailment pair in a data set and various classifier methods are evaluated based on the instance data derived from the extracted features. The focus of the paper is to compare and contrast the performance of single and ensemble based learning algorithms for a number of data sets. We showed that there is some benefit to the use of ensemble approaches but, based on the extracted features, Naïve Bayes proved to be the strongest learning mechanism. Only one ensemble approach demonstrated a slight improvement over the technique of Naïve Bayes.  相似文献   

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This publication discusses ways of using cognitive analyses for semantically interpreting economic figures. A semantic analysis making use of mathematical linguistic algorithms in order to extract meaning from sets of analysed data is illustrated with an example of a class of cognitive systems designed to analyse economic figures, or more precisely, financial ratios. The cognitive analysis systems presented in this publication are discussed as exemplified by the class of Cognitive Financial Analysis Information Systems (CFAIS). This publication proposes algorithms executed by a broad class of automatic data interpretation and understanding systems designed for the in-depth semantic analysis and interpretation of the results obtained. This will be done by defining new system classes as applications supporting decision-making processes, useful in various areas of knowledge.  相似文献   

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企业信息系统建设和集成过程产生的复杂度很大程度上决定于系统业务模型的语义复杂度。建立企业业务系统的语义复杂度测度模型是有效控制复杂度攀升的关键。建立企业系统的同构本体结构及其形式化方法的相关定义、定理,给出语义复杂度定义和测度方法。与聚合体系结构方法建模的企业系统进行语义复杂度的比较分析。结果显示,基于同构本体结构的企业系统建模过程具有最低的语义复杂度,语义复杂度不随系统建模过程发生变化,并且调整系统规模和系统集成过程不产生新的信息熵。理论上证实同构本体结构很可能是企业系统建模的最优结构,研究成果为企业系统开发和建模提供理论方法的指导。  相似文献   

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Bipolar membranes (BMs) have interesting applications within the field of bioelectronics, as they may be used to create non-linear ionic components (e.g., ion diodes and transistors), thereby extending the functionality of, otherwise linear, electrophoretic drug delivery devices. However, BM based diodes suffer from a number of limitations, such as narrow voltage operation range and/or high hysteresis. In this work, we circumvent these problems by using a novel polyphosphonium-based BM, which is shown to exhibit improved diode characteristics. We believe that this new type of BM diode will be useful for creating complex addressable ionic circuits for delivery of charged biomolecules.Combined electronic and ionic conduction makes organic electronic materials well suited for bioelectronics applications as a technological mean of translating electronic addressing signals into delivery of chemicals and ions.1 For complex regulation of functions in cells and tissues, a chemical circuit technology is necessary in order to generate complex and dynamic signal gradients with high spatiotemporal resolution. One approach to achieve a chemical circuit technology is to use bipolar membranes (BMs), which can be used to create the ionic equivalents of diodes2, 3, 4, 5 and transistors.6, 7, 8 A BM consists of a stack of a cation- and an anion-selective membrane, and functions similar to the semiconductor PN-junction, i.e., it offers ionic current rectification9, 10 (Figure ​(Figure1a).1a). The high fixed charge concentration in a BM configuration make them more suited in bioelectronic applications as compared to other non-linear ionic devices, such as diodes constructed from surface charged nanopores11 or nanochannels,12 as the latter devices typically suffers from reduced performance at elevated electrolyte concentration (i.e., at physiological conditions) due to reduced Debye screening length.13 However, a unique property of most BMs, as compared to the electronic PN-junction and other ionic diodes, is the electric field enhanced (EFE) water dissociation effect.10, 14 This occurs above a threshold reverse bias voltage, typically around −1 V, as the high electric field across the ion-depleted BM interface accelerates the forward reaction rate of the dissociation of water into H+ and OH− ions. As these ions migrate out from the BM, there will be an increase in the reverse bias current. The EFE water dissociation is a very interesting effect and is commonly used in industrial electrodialysis applications,15 where highly efficient water dissociating BMs are being researched.16 Also, BMs have also been utilized to generate H+ and OH− ions in lab-on-a-chip applications.2, 17 However, the EFE water dissociation effect diminishes the diode property of BMs when operated outside the ±1 V window, which is unwanted in, for instance, chemical circuits and addressing matrices for delivery of complex gradients of chemical species. The effect can be suppressed by incorporating a neutral electrolyte inside the BM,10, 18 for instance, poly(ethylene glycol) (PEG).2, 6, 7 However, as previously reported,2 the PEG volume will introduce hysteresis when switching from forward to reverse bias, due to its ability to store large amounts of charges. This was circumvented by ensuring that only H+ and OH− are present in the diode, which recombines into water within the PEG volume. Such diodes are well suited as integrated components in chemical circuits for pH-regulation, due to the in situ formed H+ and OH−, but are less attractive if, for instance, other ions, e.g., biomolecules, are to be processed or delivered in and from the circuit. Furthermore, a PEG electrolyte introduces additional patterning layers, making device downscaling more challenging. This is undesired when designing complex, miniaturized, and large-scale ionic circuits. Thus, there is an interest in BM diodes that intrinsically do not exhibit any EFE water dissociation, are easy to miniaturize, and that turn off at relatively high speeds. It has been suggested that tertiary amines in the BM interface are important for efficient EFE water dissociation,19, 20, 21 as they function as a weak base and can therefore catalyze dissociation of water by accepting a proton. For example, anion-selective membranes that have undergone complete methylation, converting all tertiary amines to quaternary amines, shows no EFE water dissociation,19 although this effect was not permanent, as the quaternization was reversed upon application of a current. Similar results were found for anion-selective membranes containing alkali-metal complexing crown ethers as fixed charges.21 Also, EFE water dissociation was not observed or reduced in BMs with poor ion selectivity, for example, in BMs with low fixed-charge concentration5 or with predominantly secondary and tertiary amines in the anion-selective membrane,22 as the increased co-ion transport reduces the electric field at the BM interface. However, due to decreased ion selectivity, these membranes show reduced rectification. In this work, we present a non-amine based BM diode that avoids EFE water dissociation, enables easy miniaturization, and provides fast turn-off speeds and high rectification.Open in a separate windowFigure 1(a) Ionic current rectification in a BM. In forward bias, mobile ions migrate towards the interface of the BM. The changing ion selectivity causes ion accumulation, resulting in high ion concentration and high conductivity. At high ion concentration, the selectivity of the membranes fails (Donnan exclusion failure), and ions start to pass the BM. In reverse bias, the mobile ions migrate away from the BM, eventually giving a zone with low ion concentration and low conductivity. Reverse bias can cause EFE water dissociation, producing H+ and OH- ions. (b) Chemical structures of PSS, qPVBC, and PVBPPh3. (c) The device used to characterize the BMs and the BM1A, BM2A, and BM1P designs. The BM interfaces are 50 × 50 μm.An anion-selective phosphonium-based polycation (poly(vinylbenzyl chloride) (PVBC) quaternized by triphenylphospine, PVBPPh3) was synthesized and compared to the ammonium-based polycation (PVBC quaternized by dimethylbenzylamine, qPVBC) previously used in BM diodes2 and transistors,7, 8 when included in BM diode structures together with polystyrenesulfonate (PSS) as the cation-selective material (Figure ​(Figure1b).1b). Three types of BM diodes were fabricated using standard photolithography patterning (Figure ​(Figure1c),1c), either with qPVBC (BM1A and BM2A) or PVBPPh3 (BM1P) as polycation and either with (BM2A) or without PEG (BM1A and BM1P). Poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) electrodes covered with aqueous electrolytes were used to convert electronic input signals into ionic currents through the BMs, according to the redox reaction PEDOT+:PSS− + M+ + e− ↔ PEDOT0 + M+:PSS−.The rectifying behavior of the diodes was evaluated using linear sweep voltammetry (Figure ​(Figure2).2). The BM1A diode exhibited an increase in the reverse bias current for voltages lower than −1 V, a typical signature of EFE water dissociation,10, 14 which decreased the current rectification at ±4 V to 6.14. No such deviation in the reverse bias current was observed for BM2A and BM1P, which showed rectification ratios of 751 and 196, respectively. In fact, for BM1P, no evident EFE water dissociation was observed even at −40 V (see inset of Figure ​Figure2).2). Thus, the PVBPPh3 polycation allows BM diodes to operate at voltages beyond the ±1 V window with maintained high ion current rectification.Open in a separate windowFigure 2Linear sweep voltammetry from −4 to +4 V (25 mV/s) for the BM diodes. The inset shows BM1P scanning from −40 V to +4 V (250 mV/s).The dynamic performance of the diodes was tested by applying a square wave pulse from reverse bias to a forward bias voltage of 4 V with 5–90 s pulse duration (Figure ​(Figure3).3). To access the amount of charge injected and extracted during the forward bias and subsequent turn off, the current through the device was integrated. For BM2A, we observed that the fall time increased with the duration of the forward bias pulse. This hysteresis is due to the efficient storage of ions in the large PEG volume, with no ions crossing the BM due to Donnan exclusion failure.2 As a result, during the initial period of the return to reverse bias, a large amount of charge needs to be extracted in order to deplete the BM. After a 90 s pulse, 90.6% of the injected charge during the forward bias was extracted before turn-off. This may be contrasted with BM1P, where the fall time is hardly affected by the pulse duration, and the extracted/injected ratio is only 15.4% for a 90 s pulse. For this type of BM, the interface quickly becomes saturated with ions during forward bias, leading to Donnan exclusion failure and transport of ions across the BM.4 Thus, less charge needs to be extracted to deplete the BM, allowing for faster fall times and significantly reduced hysteresis.Open in a separate windowFigure 3Switching characteristics (5, 10, 20, 30, 60, or 90 s pulse) and ion accumulation (at 90 s pulse) of the BM2A and BM1P diodes. BM1A showed similar characteristics as BM1P when switched at ±1V (see supplementary material).24Since the neutral electrolyte is no longer required to obtain high ion current rectification over a wide potential range, the size of the BM can be miniaturized. This translates into higher component density when integrating the BM diode into ionic/chemical circuits. A miniaturized BM1P diode was constructed, where the interface of the BM was shrunk from 50 μm to 10 μm. The 10 μm device showed similar IV and switching characteristics as before (Figure ​(Figure4),4), but with higher ion current rectification ratio (over 800) and decreased rise/fall times (corresponding to 90%/–10% of forward bias steady state) from 10 s/12.5 s to 4 s/4 s. Since the overlap area is smaller, a probable reason for the faster switching times is the reduced amount of ions needed to saturate and deplete the BM interface. In comparison to our previous reported low hysteresis BM diode,2 this miniaturized polyphosphonium-based devices shows the same rise and fall times but increased rectification ratio.Open in a separate windowFigure 4(a) Linear sweep voltammetry and (b) switching performance of a BM1P diode with narrow junction.In summary, by using polyphosphonium instead of polyammonium as the polycation in BM ion diodes the EFE water dissociation can be entirely suppressed over a large operational voltage window, supporting the theory that a weak base, such as a tertiary amine, is needed for efficient EFE water dissociation.17, 18 As no additional neutral layer at the BM interface is needed, ion diodes that operate outside the usual EFE water dissociation window of ±1 V can be constructed using less active layers, fewer processing steps and with relaxed alignment requirement as compared to polyammonium-based devices. This enables the fabrication of ion rectification devices with an active interface as low as 10 μm. Furthermore, the exclusion of a neutral layer improves the overall dynamic performance of the BM ion diode significantly, as there is less hysteresis due to ion accumulation. Previously, the hysteresis of BM ion diodes has been mitigated by designing the diode so that only H+ and OH− enters the BM, which then recombines into water.2 Such diodes also show high ion current rectification ratio and switching speed but are more complex to manufacture, and their application in organic bioelectronic systems is limited due to the H+/OH− production. By instead using the polyphosphonium-based BM diode, reported here, we foresee ionic, complex, and miniaturized circuits that can include charged biomolecules as the signal carrier to regulate functions and the physiology in cell systems, such as in biomolecule and drug delivery applications, and also in lab-on-a-chip applications.  相似文献   

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A flow redirection and single cell immobilization method in a microfluidic chip is presented. Microheaters generated localized heating and induced poly(N-isopropylacrylamide) phase transition, creating a hydrogel that blocked a channel or immobilized a single cell. The heaters were activated in sets to redirect flow and exchange the fluid in which an immobilized cell was immersed. A yeast cell was immobilized in hydrogel and a 4′,6-diamidino-2-phenylindole (DAPI) fluorescent stain was introduced using flow redirection. DAPI diffused through the hydrogel and fluorescently labelled the yeast DNA, demonstrating in situ single cell biochemistry by means of immobilization and fluid exchange.The ability to control microfluidic flow is central to nearly all lab-on-a-chip processes. Recent developments in microfluidics either include microchannel based flow control in which microvalves are used to control the passage of fluid,1 or are based on discrete droplet translocation in which electric fields or thermal gradients are used to determine the droplet path.2, 3 Reconfigurable microfluidic systems have certain advantages, including the ability to adapt downstream fluid processes such as sorting to upstream conditions and events. This is especially relevant for work with individual biomolecules and high throughput cell sorting.4 Additionally, reconfigurable microfluidic systems allow for rerouting flows around defective areas for high device yield or lifetime and for increasing the device versatility as a single chip design can have a variety of applications.Microvalves often form the basis of flow control systems and use magnetic, electric, piezoelectric, and pneumatic actuation methods.5 Many of these designs require complicated fabrication steps and can have large complex structures that limit the scalability or feasability of complex microfluidic systems. Recent work has shown how phase transition of stimuli-responsive hydrogels can be used to actuate a simple valve design.6 Beebe et al. demonstrated pH actuated hydrogel valves.7 Phase transition of thermosensitive poly(N-isopropylacrylamide) (PNIPAAm) using a heater element was demonstrated by Richter et al.8 Phase transition was also achieved by using light actuation by Chen et al.9 Electric heating has shown a microflow response time of less than 33 ms.11 Previous work10 showed the use of microheaters to induce a significant shift in the viscosity of thermosensitive hydrogel to block microchannel flow and deflect a membrane, stopping flow in another microchannel. Additionally, Yu et al.12 demonstrated thermally actuated valves based on porous polymer monoliths with PNIPAAm. Krishnan and Erickson13 showed how reconfigurable optically actuated hydrogel formation can be used to dynamically create highly viscous areas and thus redirect flow with a response time of  ～ 2?s. This process can be used to embed individual biomolecules in hydrogel and suppress diffusion as also demonstrated by others.15, 16 Fiddes et al.14 demonstrated the use of hydrogels to transport immobilized biomolecules in a digital microfluidic system. While the design of Krishnan and Erickson is highly flexible, it requires the use of an optical system and absorption layer to generate a geometric pattern to redirect flow.This paper describes the use of an array of gold microheaters positioned in a single layer polydimethylsiloxane (PDMS) microfluidic network to dynamically control microchannel flow of PNIPAAm solution. Heat generation and thus PNIPAAm phase transition were localized as the microheaters were actuated using pulse width modulation (PWM) of an applied electric potential. Additionally, hydrogel was used to embed and immobilise individual cells, exchange the fluid parts of the microfluidic system in order to expose the cells to particular reagents to carry out an in situ biochemical process. The PDMS microchannel network and the microheater array are shown in Figure ​Figure11.Open in a separate windowFigure 1A sketch of the electrical circuit and a microscope image of the gold microheaters and the PDMS microchannels. The power to the heaters was modulated with a PWM input through a H-bridge. For clarity, the electrical circuit for only the two heaters with gelled PNIPAAm is shown (H1 and V2). There are four heaters (V1-V4) in the “vertical channels” and three heaters (H1-H3) in the “horizontal” channel.The microchannels were fabricated using a patterned mould on a silicon wafer to define PDMS microchannels, as described by DeBusschere et al.17 and based on previous work.10 A 25 × 75 mm glass microscope slide served as the remaining wall of the microchannel system as well as the substrate for the microheater array. The gold layer had a thickness of 200 nm and was deposited and patterned using E-beam evaporation and photoresist lift-off.21 The gold was patterned to function as connecting electrical conductors as well as the microheaters.It was crucial that the microheater array was aligned with an accuracy of  ～ 20μm with the PDMS microchannel network for good heat localization. The PDMS and glass lid were treated with plasma to activate the surface and alignment was carried out by mounting the microscope slide onto the condenser lens of an inverted microscope (TE-2000 Nikon Instruments). While imaging with a 4× objective, the x, y motorized stage aligned the microchannels to the heaters and the condenser lens was lowered for the glass substrate to contact the PDMS and seal the microchannels.Local phase transition of 10% w/w PNIPAAm solution in the microchannels was achieved by applying a 7 V potential through a H-bridge that received a PWM input at 500 Hz which was modulated using a USB controller (Arduino Mega 2650) and a matlab (Mathworks) GUI. The duty cycle of the PWM input was calibrated for each microheater to account for differences in heater resistances (25?Ω to 52?Ω) due to varying lengths of on-chip connections and slight fabrication inconsistencies, as well as for different flow conditions during device operation. Additionally, thermal cross-talk between heaters required decreasing the PWM input significantly when multiple heaters were activated simultaneously. This allowed confining the areas of cross-linked PNIPAAm to the microheaters, allowing the fluid in other areas to flow freely.By activating the heaters in sets, it was possible to redirect the flow and exchange the fluid in the central area. Figure ​Figure22 demonstrates how the flow direction in the central microchannel area was changed from a stable horizontal flow to a stable vertical flow with a 3 s response time, using only PNIPAAm phase transition. Constant pressures were applied to the inlets to the horizontal channel and to the vertical channels. Activating heaters V1-4 (Figure ​(Figure2,2, left) resulted in flow in the horizontal channel only. Likewise, activating heaters H1 and H2 allowed for flow in the vertical channel only. In this sequence, the fluid in the central microchannel area from one inlet was exchanged with fluid from the other inlet. Additionally, by activating heater H3, a particle could be immobilised during the exchange of fluid as shown in Figure ​Figure33 (top).Open in a separate windowFigure 2Switching between fluid from the horizontal and the vertical channel using hydrogel activation and flow redirection with a response time of 3 s. A pressure of 25 mbar was applied to the inlet of the horizontal channel and a pressure of 20 mbar to the vertical channel. The flow field was determined using particle image velocimetry, in which the displacement of fluorescent seed particles was determined from image pairs generated by laser pulse exposure. Processing was carried out with davis software (LaVision).Open in a separate windowFigure 3A series of microscope images near heater H3 showing: (1a)-(1c) A single yeast cell captured by local PNIPAAm phase transition and immobilized for 5 min before being released. (2a) A single yeast cell was identified for capture by embedding in hydrogel. (2b) The cell as well as the hydrogel displayed fluorescence while embedded due to the introduction of DAPI in the surrounding region. (2c) The diffusion of DAPI towards the cell as the heating power of H3 is reduced after 15 min, showing a DAPI stained yeast cell immobilized.Particle immobilisation in hydrogel and fluid exchange in the central area of the microfluidic network were used to carry out an in situ biochemical process in which a yeast cell injected through one inlet was stained in situ with a 4′,6-diamidino-2-phenylindole (DAPI) solution (Invitrogen), which attached to the DNA of the yeast cell.18 A solution of yeast cells with a concentration of 5 × 107cells/ml suspended in a 10% w/w PNIPAAm solution was injected through the horizontal channel. A solution of 2μg/l DAPI in a 10% w/w PNIPAAm solution was injected through the vertical channel. A single yeast cell was identified and captured near the central heater, and by deactivating the heaters in the vertical channel, DAPI solution was introduced in the microchannels around the hydrogel. After immobilising the cell for 15 min, the heater was deactivated, releasing the cell in the DAPI solution. This process is shown in Figure ​Figure33 (bottom). The sequence of the heater activation and deactivation in order to immobilize the cell and exchange the fluid is outlined in the supplementary material.21Eriksen et al.15 demonstrated the diffusion of protease K in the porous hydrogel matrix,19 and it was therefore expected that DAPI fluorescent stain (molecular weight of 350 kDa, Ref. 20) would also diffuse. DAPI diffusion is shown in Figure 3(2b) in which the yeast cell shows fluorescence while embedded in the hydrogel. The yeast cell was released by deactivating the central heater and activating all the others to suppress unwanted flow in the microchannel. As a result, the single cell was fully immersed in the DAPI solution. Immobilization of a single cell allows for selection of a cell that exhibits a certain trait and introduction of a new fluid while maintaining the cell position in the field of view of the microscope such that a biochemical response can be imaged continuously.In summary, a microfluidic chip capable of local heating was used to induce phase transition of PNIPAAm to hydrogel, blocking microchannel flow, and thereby allowing for reconfigurable flow. Additionally, the hydrogel was used to embed and immobilise a single yeast cell. DAPI fluorescent stain was introduced using flow redirection, and it stained the immobilized cell, showing diffusion into the hydrogel. The versatile design of this microfluidic chip permits flow redirection, and is suitable to carry out in situ biochemical reactions on individual cells, demonstrating the potential of this technology for forming large-scale reconfigurable microfluidic networks for biochemical applications.  相似文献   

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