Point-of-Care Diagnostic Devices for Detection of Escherichia coli O157:H7 Using Microfluidic Systems: A Focused Review.

Bacterial infections represent a serious and global threat in modern medicine; thus, it is very important to rapidly detect pathogenic bacteria, such as Escherichia coli (E. coli) O157:H7. Once treatments are delayed after the commencement of symptoms, the patient's health quickly deteriorates. Hence, real-time detection and monitoring of infectious agents are highly critical in early diagnosis for correct treatment and safeguarding public health. To detect these pathogenic bacteria, many approaches have been applied by the biosensors community, for example, widely-used polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), culture-based method, and adenosine triphosphate (ATP) bioluminescence. However, these approaches have drawbacks, such as time-consumption, expensive equipment, and being labor-intensive, making it critical to develop ultra-sensitive and highly selective detection. The microfluidic platform based on surface plasmon resonance (SPR), electrochemical sensing, and rolling circle amplification (RCA) offers proper alternatives capable of supplementing the technological gap for pathogen detection. Note that the microfluidic biochip allows to develop rapid, sensitive, portable, and point-of-care (POC) diagnostic tools. This review focuses on recent studies regarding accurate and rapid detection of E. coli O157:H7, with an emphasis on POC methods and devices that complement microfluidic systems. We also examine the efficient whole-body detection by employing antimicrobial peptides (AMPs), which has attracted growing attention in many applications.

Impaired migration of autologous induced neural stem cells from patients with schizophrenia and implications for genetic risk for psychosis.

Stem cell technologies have presented explicit evidence of the neurodevelopmental hypothesis of schizophrenia. However, few studies investigated relevance of the schizophrenia genetic liability and the use of genetic reprogramming on pluripotent stem cells to the impaired neurodevelopment shown by stem cells. Therefore, this study sought to investigate the cellular phenotypes of induced neural stem cells (iNSCs) derived without genetic modification from patients with schizophrenia and from genetic high risk (GHR) individuals. Three patients with a diagnosis of schizophrenia, 3 GHR individuals who had two or more relatives with schizophrenia, and 3 healthy volunteers participated. iNSCs were derived using a small molecule-based lineage switch method, and their gene expression levels and migration capabilities were examined. Demographic characteristics were not different among the groups (age, χ2 = 5.637, P = .060; education, χ2 = 2.111, P = .348). All participants stayed well during the follow-up except one GHR individual who developed psychosis 1.5 years later. Migration capacity was impaired in iNSCs from patients with schizophrenia (SZ-iNSCs) compared to iNSCs from GHR individuals or controls (P < .001). iNSCs from a GHR individual who later developed schizophrenia showed migratory impairment that was similar to SZ-iNSCs. Gene expression levels of Sox2 in SZ-iNSCs were significantly lower than those in controls (P = .028). Defective migration in genetically unmodified SZ-iNSCs is the first direct demonstration of neurodevelopmental abnormalities in schizophrenia. Additionally, alterations in gene expression in SZ-iNSCs suggest mechanisms by which genetic liability leads to aberrant neurodevelopment.

Electrostatic Potential Analysis in Polyelectrolyte Brush-Grafted Microchannels Filled with Polyelectrolyte Dispersion.

In this study, the model framework that includes almost all relevant parameters of interest has been developed to quantify the electrostatic potential and charge density occurring in microchannels grafted with polyelectrolyte brushes and simultaneously filled with polyelectrolyte dispersion. The brush layer is described by the Alexander-de Gennes model incorporated with the monomer distribution function accompanying the quadratic decay. Each ion concentration due to mobile charges in the bulk and fixed charges in the brush layer can be determined by multi-species ion balance. We solved 2-dimensional Poisson-Nernst-Planck equations adopted for simulating electric field with ion transport in the soft channel, by considering anionic polyelectrolyte of polyacrylic acid (PAA). Remarkable results were obtained regarding the brush height, ionization, electrostatic potential, and charge density profiles with conditions of brush, dispersion, and solution pH. The Donnan potential in the brush channel shows several times higher than the surface potential in the bare channel, whereas it becomes lower with increasing PAA concentration. Our framework is fruitful to provide comparative information regarding electrostatic interaction properties, serving as an important bridge between modeling and experiments, and is possible to couple with governing equations for flow field.

Two-phase flow in microfluidic-chip design of hydrodynamic filtration for cell particle sorting.

As one of the flow-based passive sorting, the hydrodynamic filtration using a microfluidic-chip has shown to effectively separate into different sizes of subpopulations from cell or particle suspensions. Its model framework involving two-phase Newtonian or generalized Newtonian fluid (GNF) was developed, by performing the complete analysis of laminar flow and complicated networks of main and multiple branch channels. To predict rigorously what occurs in flow fields, we estimated pressure drop, velocity profile, and the ratio of the flow fraction at each branch point, in which the analytical model was validated with numerical flow simulations. As a model fluid of the GNF, polysaccharide solution based on Carreau type was examined. The objective parameters aiming practical channel design include the number of the branches and the length of narrow section of each branch for arbitrary conditions. The flow fraction and the number of branches are distinctly affected by the viscosity ratio between feed and side flows. As the side flow becomes more viscous, the flow fraction increases but the number of branches decreases, which enables a compact chip designed with fewer branches being operated under the same throughput. Hence, our rational design analysis indicates the significance of constitutive properties of each stream.

Efficient detection of Escherichia coli O157:H7 using a reusable microfluidic chip embedded with antimicrobial peptide-labeled beads.

The ability of antimicrobial peptides (AMPs) for effective binding to multiple target microbes has drawn lots of attention as an alternative to antibodies for detecting whole bacteria. We investigated pathogenic Escherichia coli (E. coli) detection by applying a microfluidic based biosensing device embedded with AMP-labeled beads. According to a new channel design, our device is reusable by the repeated operation of detection and regeneration modes, and the binding rate is more enhanced due to even distribution of the bacterial suspension inside the chamber by implementing influx side channels. We observed higher binding affinity of pathogenic E. coli O157:H7 for AMP-labeled beads than nonpathogenic E. coli DH5α, and the fluorescence intensity of pathogenic E. coli was about 3.4 times higher than the nonpathogenic one. The flow rate of bacterial suspension should be applied above a certain level for stronger binding and rapid detection by attaining a saturation level of detection within a short time of less than 20 min. A possible improvement in the limit of detection in the level of 10 cells per mL for E. coli O157:H7 implies that the AMP-labeled beads have high potential for the sensitive detection of pathogenic E. coli at an appropriate flow rate.

Sorting of human mesenchymal stem cells by applying optimally designed microfluidic chip filtration.

Human bone marrow-derived mesenchymal stem cells (hMSCs) consist of heterogeneous subpopulations with different multipotent properties: small and large cells with high and low multipotency, respectively. Accordingly, sorting out a target subpopulation from the others is very important to increase the effectiveness of cell-based therapy. We performed flow-based sorting of hMSCs by using optimally designed microfluidic chips based on the hydrodynamic filtration (HDF) principle. The chip was designed with the parameters rigorously determined by the complete analysis of laminar flow for flow fraction and complicated networks of main and multi-branched channels for hMSCs sorting into three subpopulations: small (<25 µm), medium (25-40 µm), and large (>40 µm) cells. By focusing with a proper ratio between main and side flows, cells migrate toward the sidewall due to a virtual boundary of fluid layers and enter the branch channels. This opens the possibility of sorting stem cells rapidly without damage. Over 86% recovery was achieved for each population of cells with complete purity in small cells, but the sorting efficiency of cells is slightly lower than that of rigid model particles, due to the effect of cell deformation. Finally, we confirmed that our method could successfully fractionate the three subpopulations of hMSCs by analyzing the surface marker expressions of cells from each outlet.

Electrokinetic secondary-flow behavior in a curved microchannel under dissimilar surface conditions.

The curved channel appears to be indispensable for the lab-on-chips systems because it provides a convenient scheme for increasing the channel length per unit chip area in the direction of net flow. A secondary Dean flow in curved rectangular microchannels is examined by applying the finite-volume scheme with a semi-implicit method for pressure-linked equations (SIMPLE) algorithm for the pressure-driven electrokinetic transport. This framework is based on the theoretical model coupled with the full Poisson-Boltzmann, Navier-Stokes, and the Nernst-Planck principles of net charge conservation [Yun et al., Phys. Fluids 22, 052004 (2010)]. The effect of a dissimilar wall condition on the secondary flow at the turn is explored by considering different configurations of channel wall having complementary aspect ratios (i.e., ratio of the channel height to the channel width, H/W = 0.25 and 4.0) with combinations of hydrophilic glass and hydrophobic polydimethylsiloxane surfaces. Simulation results exhibit that, contrary to the case of general narrow-bore channels, the streamwise axial velocity tends to shift toward the inner wall caused by a stronger effect of the spanwise pressure gradient, according to a sufficiently low Dean number. The increasing rate of this shift with increasing curvature ratio is more significant in the shallow (or low-aspect-ratio) channel, due to the effect of greater distance traveled by the fluid along the outer wall. The curvature introduces the presence of pairs of counter-rotating vortices perpendicular to the flow direction. Comparing between shallow and deep (or high-aspect-ratio) channels allows us to identify that the patterns of axial velocity and vorticity are altered by the heterogeneity effect of surfaces occupying a large area. The total magnitude of vorticity at the cross section of the channel increases with increasing slip length, due to the contribution of enhanced axial velocity driven by the slip, while there is no fluid-slip dependency for the slip length of less than about 50 nm.

Conformation and translational diffusion of a xanthan polyelectrolyte chain: Brownian dynamics simulation and single molecule tracking.

In our recent Brownian dynamics (BD) simulation study, the structure and dynamics of anionic polyelectrolyte xanthan in bulk solution as well as confined spaces of slitlike channel were examined by applying a coarse-grained model with nonlinear bead-spring discretization of a whole chain [J. Jeon and M.-S. Chun, J. Chem. Phys. 126, 154904 (2007)]. This model goes beyond other simulations as they did not consider both long-range electrostatic and hydrodynamic interactions between pairs of beads. Simulation parameters are obtained from the viscometric method of rheology data on the native and sonicated xanthan polysaccharides, which have a contour length less than 1 microm . The size of the semiflexible polyelectrolyte can be well described by the wormlike chain model once the electrostatic effects are taken into account by the persistence length measured at a long length scale. For experimental verifications, single molecule visualization was performed on fluorescein-labeled xanthan using an inverted fluorescence microscope, and the motion of an individual molecule was quantified. Experimental results on the conformational changes in xanthan chain in the electrolyte solution have a reasonable trend to agree with the prediction by BD simulations. In the translational diffusion induced by the Debye screening effect, the simulation prediction reveals slightly higher values compared to those of our measurements, although it agrees with the literature data. Considering the experimental restrictions, our BD simulations are verified to model the single polyelectrolyte well.

Structure of flexible and semiflexible polyelectrolyte chains in confined spaces of slit micro/nanochannels.

Understanding the behavior of a polyelectrolyte in confined spaces has direct relevance in design and manipulation of microfluidic devices, as well as transport in living organisms. In this paper, a coarse-grained model of anionic semiflexible polyelectrolyte is applied, and its structure and dynamics are fully examined with Brownian dynamics (BD) simulations both in bulk solution and under confinement between two negatively charged parallel plates. The modeling is based on the nonlinear bead-spring discretization of a continuous chain with additional long-range electrostatic, Lennard-Jones, and hydrodynamic interactions between pairs of beads. The authors also consider the steric and electrostatic interactions between the bead and the confining wall. Relevant model parameters are determined from experimental rheology data on the anionic polysaccharide xanthan reported previously. For comparison, both flexible and semiflexible models are developed accompanying zero and finite intrinsic persistence lengths, respectively. The conformational changes of the polyelectrolyte chain induced by confinements and their dependence on the screening effect of the electrolyte solution are faithfully characterized with BD simulations. Depending on the intrinsic rigidity and the medium ionic strength, the polyelectrolyte can be classified as flexible, semiflexible, or rigid. Confined flexible and semiflexible chains exhibit a nonmonotonic variation in size, as measured by the radius of gyration and end-to-end distance, with changing slit width. For the semiflexible chain, this is coupled to the variations in long-range bond vector correlation. The rigid chain, realized at low ionic strength, does not have minima in size but exhibits a sigmoidal transition. The size of confined semiflexible and rigid polyelectrolytes can be well described by the wormlike chain model once the electrostatic effects are taken into account by the persistence length measured at long length scale.

Surface properties of submicrometer silica spheres modified with aminopropyltriethoxysilane and phenyltriethoxysilane.

The surface of submicrometer silica spheres are modified with aminopropyl and phenyl groups through a one-step process. Various experimental techniques, i.e., scanning electron microscopy (SEM), quasi-elastic light scattering (QELS), differential scanning calorimetry (DSC), thermogravimetry (TG), zeta potential measurement, nitrogen sorption, and water vapor and organic dye adsorption are used to comprehensively characterize the pure (TEOS particles) and modified silica particles. The SEM micrographs of the particles demonstrate that the modified particles are spherical with uniform size and shape. The particles modified with aminopropyl groups (APTES particles) show the highest isoelectric point (IEP) and the highest weight loss at 780 degrees C because of the basic nature of aminopropyl groups and the higher reactivity of aminopropyltriethoxysilane. The particles modified with the phenyl groups (PhTES particles) show the lowest water vapor adsorption because their surface is more hydrophobic than that of TEOS and APTES particles. The organic dye (brilliant blue FCF or BBF) adsorption experiments demonstrate that the adsorption capacity of the particles increases greatly after acidification. This is caused by the protonation of silanol groups and amine groups on the particle surface, which presents an enhanced electrostatic attraction with BBF anions. The APTES particles exhibit the highest dye adsorption due to the hydrophobic attractions and the enhanced electrostatic attractions from aminopropyl groups.

Fabrication and validation of a multi-channel type microfluidic chip for electrokinetic streaming potential devices.

To elaborate on the applicability of the electrokinetic micro power generation, we designed and fabricated the silicon-glass as well as the PDMS-glass microfluidic chips with the unique features of a multi-channel. Besides miniaturizing the device, the key advantage of our microfluidic chip utilization lies in the reduction in water flow rate. Both a distributor and a collector taking the tapered duct geometry are positioned aiming the uniform distribution of water flow into all individual channels of the chip, in which several hundreds of single microchannels are assembled in parallel. A proper methodology is developed accompanying the deep reactive ion etching as well as the anodic bonding, and optimum process conditions necessary for hard and soft micromachining are presented. It has been shown experimentally and theoretically that the silicon-based microchannel leads to increasing streaming potential and higher external current compared to those of the PDMS-based one. A proper comparison between experimental results and theoretical computations allows justification of the validity of our novel devices. It is useful to recognize that a material inducing a higher magnitude of zeta potential has an advantage for obtaining higher power density under the same external resistance.

Surface functional group effect on atomic force microscope anodization lithography.

The lithographic effect of surface chemical functional groups of organic resists on atomic force microscope (AFM) anodization lithography is investigated using mixed self-assembled monolayers (SAMs). The SAM resist films were prepared with 1,12-diaminododecane dihydrochloride (DAD.2HCl), n-tridecylamine hydrochloride (TDA.HCl), and 1,12-diaminododecane hydrochloride (DAD.HCl), and their film characteristics were evaluated by ellipsometry, zeta-potential measurements, and AFM. The lithographic results indicate that the most dominant factor of the surface functional group effect is the electrochemical property of the surface groups as an anode surface in the anodization reaction, and the dimensions of the protruded patterns are critically determined by the wetting property of the resist surface. By controlling the surface chemical groups with considerations of their effects, high-speed patterning at 2 mm/s was achieved successfully using the mixed SAM resist of DAD.2HCl and TDA.HCl.

Rigorous calculations of linearized Poisson-Boltzmann interaction between dissimilar spherical colloids and osmotic pressure in concentrated dispersions.

We present computational results on the static properties of concentrated dispersions of bidisperse colloids. The long-range electrostatic interactions between dissimilar spherical colloids are determined using the singularity method, which provides rigorous solutions to the linearized electrostatic field. The NVT Monte Carlo simulation is applied to the bulk suspension to obtain the radial distribution function for the concentrated system. The increasing trend of osmotic pressure with increasing total particle concentration is reduced as the concentration ratio between large and small particles is increased. The increase of electrostatic interaction between similarly charged particles caused by the Debye screening effect provides an increase in the osmotic pressure. From the estimation of total structure factor, we observe the strong correlations developed between dissimilar spheres, and the small spheres are expected to tend to fit into the spaces between the larger ones. As the particle concentration increases at a given ionic strength, the magnitude of the first peak in structure factors increases and also moves to higher wavenumber values.

Estimation of zeta potential by electrokinetic analysis of ionic fluid flows through a divergent microchannel.

The streaming potential is generated by the electrokinetic flow effect within the electrical double layer of a charged solid surface. Surface charge properties are commonly quantified in terms of the zeta potential obtained by computation with the Helmholtz-Smoluchowski (H-S) equation following experimental measurement of streaming potential. In order to estimate a rigorous zeta potential for cone-shaped microchannel, the correct H-S equation is derived by applying the Debye-Hückel approximation and the fluid velocity of diverging flow on the specified position. The present computation provides a correction ratio relative to the H-S equation for straight cylindrical channel and enables us to interpret the effects of the channel geometry and the electrostatic interaction. The correction ratio decreases with increasing of diverging angle, which implies that smaller zeta potential is generated for larger diverging angle. The increase of Debye length also reduces the correction ratio due to the overlapping of the Debye length inside of the channel. It is evident that as the diverging angle of the channel goes to nearly zero, the correction ratio converges to the previous results for straight cylindrical channel.

On the behavior of electrokinetic streaming potential during protein filtration with fully and partially retentive nanopores.

An experimental investigation of the electrokinetic streaming potentials of both fully and partially retentive nanopores as compared with the filtration progress of dilute globular protein solution under different surface charge conditions was performed using hollow fibers. The streaming potential is generated by the electrokinetic flow effect within the electric double layer of the charged surface. Depending on the solution pH, both the protein and the pore wall can be either repulsive or attractive due to the long-range electrostatic interaction. The repulsive electrostatic interaction allows the protein particles to stay in a suspended state above the outer surface of hollow fibers instead of being deposited. The apparent streaming potential value at partially retentive pores is larger than that at fully retentive pores for the oppositely charged case; however, the opposite behavior is shown for the same-charged case. The axial-position-dependent streaming potential was also observed in order to explore the development of a concentration polarization layer during the cross-flow filtration. The time evolution of the streaming potential during the filtration of protein particles is related to the filtrate flux, from which it can be found to provide useful real-time information on particle deposition onto the outer surfaces of hollow fibers.