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Mar 2010

Volume 4, Issue 1, Articles (01xxxx)

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Bio-electrospraying and aerodynamically assisted bio-jetting whole human blood: Interrogating cell surface marker integrity

Pascal Joly, Naina Chavda, Ayad Eddaoudi, and Suwan N. Jayasinghe

Biomicrofluidics 4, 011101 (2010); http://dx.doi.org/10.1063/1.3294083 (6 pages) | Cited 4 times

Online Publication Date: 13 January 2010

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Bio-electrospraying and aerodynamically assisted bio-jetting are two direct cell handling approaches recently pioneered, which have demonstrated significant applicability to the life sciences. These two bioprotocols have undergone scientific rigor, which have seen these techniques been explored in conjunction with a wide range of immortalized, primary and stem cells, and those whole organisms. Those studies have demonstrated a cellular population of >70% viable post-treatment in comparison with controls. Although, these studies assessed cellular viability, cell surface molecules play a critical role in several cellular functions, in particular, have importance to tissue engineering and regenerative medicine. Thus, in the studies reported herein, we demonstrate post-treated viable cells retain their cell surface marker expression levels in comparison to controls, over both short and long time points. Therefore, these studies further push back the frontiers of both bio-electrosprays and aerodynamically assisted bio-jetting in their endeavor as novel strategies for tissue engineering and regenerative biology/medicine with possible targeted clinical utility.
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87.80.-y Biophysical techniques (research methods)
87.85.Lf Tissue engineering
87.17.-d Cell processes

Enhancement of biosensing performance in a droplet-based bioreactor by in situ microstreaming

Olivier Ducloux, Elisabeth Galopin, Farzam Zoueshtiagh, Alain Merlen, and Vincent Thomy

Biomicrofluidics 4, 011102 (2010); http://dx.doi.org/10.1063/1.3310930 (5 pages) | Cited 1 time

Online Publication Date: 8 February 2010

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A droplet-based micro-total-analysis system involving biosensor performance enhancement by integrated surface-acoustic-wave (SAW) microstreaming is shown. The bioreactor consists of an encapsulated droplet with a biosensor on its periphery, with in situ streaming induced by SAW. This paper highlights the characterization by particle image tracking of the speed distribution inside the droplet. The analyte-biosensor interaction is then evaluated by finite element simulation with different streaming conditions. Calculation of the biosensing enhancement shows an optimum in the biosensor response. These results confirm that the evaluation of the Damköhler and Peclet numbers is of primary importance when designing biosensors enhanced by streaming.
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87.80.Ek Mechanical and micromechanical techniques
43.38.Rh Surface acoustic wave transducers
43.25.Nm Acoustic streaming
47.85.Np Fluidics
82.80.-d Chemical analysis and related physical methods of analysis
85.85.+j Micro- and nano-electromechanical systems (MEMS/NEMS) and devices

Analyzing shear stress-induced alignment of actin filaments in endothelial cells with a microfluidic assay

A. D. van der Meer, A. A. Poot, J. Feijen, and I. Vermes

Biomicrofluidics 4, 011103 (2010); http://dx.doi.org/10.1063/1.3366720 (5 pages) | Cited 3 times

Online Publication Date: 15 March 2010

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The physiology of vascular endothelial cells is strongly affected by fluid shear stress on their surface. In this study, a microfluidic assay was employed to analyze the alignment of actin filaments in endothelial cells in response to shear stress. When cells were cultured in microfluidic channels and subjected to shear stress, the alignment of filaments in the channel direction was significantly higher than in static cultures. By adding inhibitory drugs, the roles of several signaling proteins in the process of alignment were determined. Thus, it is shown how microfluidic technology can be employed to provide a mechanistic insight into cell physiology.
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85.85.+j Micro- and nano-electromechanical systems (MEMS/NEMS) and devices
87.80.Ek Mechanical and micromechanical techniques
87.17.-d Cell processes
87.19.U- Hemodynamics
07.10.Cm Micromechanical devices and systems
87.80.-y Biophysical techniques (research methods)
87.64.-t Spectroscopic and microscopic techniques in biophysics and medical physics
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Preface to Special Topic: Papers from the 13th International Conference on Surface and Colloid Science (ICSCS) and the 83rd ACS Colloid and Surface Science Symposium, Columbia University, New York, 2009

Leslie Y. Yeo

Biomicrofluidics 4, 013101 (2010); http://dx.doi.org/10.1063/1.3332379 (2 pages)

Online Publication Date: 5 March 2010

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This Special Topic section is a compilation of several original contributions covering both fundamental and practical aspects of electrokinetic microfluidic phenomena that were presented during the Electrokinetics and Microfluidics sessions held at the conference.
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01.30.-y Physics literature and publications

Dielectrophoretic choking phenomenon in a converging-diverging microchannel

Ye Ai, Shizhi Qian, Sheng Liu, and Sang W. Joo

Biomicrofluidics 4, 013201 (2010); http://dx.doi.org/10.1063/1.3279787 (6 pages) | Cited 11 times

Online Publication Date: 7 January 2010

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Experiments show that particles smaller than the throat size of converging-diverging microchannels can sometimes be trapped near the throat. This critical phenomenon is associated with the negative dc dielectrophoresis arising from nonuniform electric fields in the microchannels. A finite-element model, accounting for the particle-fluid-electric field interactions, is employed to investigate the conditions for this dielectrophoretic (DEP) choking in a converging-diverging microchannel for the first time. It is shown quantitatively that the DEP choking occurs for high nonuniformity of electric fields, high ratio of particle size to throat size, and high ratio of particle’s zeta potential to that of microchannel.
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87.80.Ek Mechanical and micromechanical techniques
47.85.Np Fluidics
82.45.-h Electrochemistry and electrophoresis
02.70.Dh Finite-element and Galerkin methods

Frequency-dependent behaviors of individual microscopic particles in an optically induced dielectrophoresis device

Xiaolu Zhu, Hong Yi, and Zhonghua Ni

Biomicrofluidics 4, 013202 (2010); http://dx.doi.org/10.1063/1.3279788 (14 pages) | Cited 5 times

Online Publication Date: 7 January 2010

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An optoelectronic microdevice is set up to drive single microparticles and a maximum synchronous velocity (MS-velocity) spectrum method is proposed for quantifying the frequency-dependent behaviors of individual neutral microparticles from 40 kHz to 10 MHz. Dielectrophoretic behaviors of three types of microparticles are investigated under the optically induced nonuniform electric field. Different MS-velocity spectra for the three different particles are experimentally found. Numerical calculations for the MS-velocity spectra of polystyrene microparticles are performed. The spectrum of the MS-velocities for a specific particle is mainly determined by the particle inherent property and the electric characteristics of the device. Moreover the experimental and the numerical MS-velocity spectra are compared to be accordant. Based on the dielectrophoretic (DEP) behaviors of the particles under a nonuniform electric field, microparticles can be finely characterized or distinguished according to their distinct MS-velocity spectra.
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87.80.Ek Mechanical and micromechanical techniques
87.80.Fe Micromanipulation of biological structures
85.85.+j Micro- and nano-electromechanical systems (MEMS/NEMS) and devices
87.50.ch Electrophoresis/dielectrophoresis and other mechanical effects
82.80.Dx Analytical methods involving electronic spectroscopy

Onset of channeling during DNA electrophoresis in a sparse ordered post array

Jia Ou, Samuel J. Carpenter, and Kevin D. Dorfman

Biomicrofluidics 4, 013203 (2010); http://dx.doi.org/10.1063/1.3283903 (10 pages) | Cited 9 times

Online Publication Date: 7 January 2010

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The “channeling hypothesis” of DNA electrophoresis in sparse, ordered arrays of posts predicts that the DNA will move through the array relatively unhindered if (i) the spacing between the posts is larger than the DNA coil and (ii) the electric field lines are straight. We tested this hypothesis by studying the electrophoretic separation of a small plasmid DNA (pUC19, 2686 base pairs) and a large, linear DNA (λ-DNA, 48 500 base pairs) in a hexagonal array of 1 μm diameter posts with a pitch of 7 μm. At low electric field strengths, these DNAs are separated due to the long-lived, rope-over-pulley collisions of λ-DNA with the posts. The resolution is lost as the electric field increases due to the onset of channeling by the λ-DNA. Using a diffusive model, we show that channeling arises at low electric fields due to the finite size of the array. This channeling is not intrinsic to the system and is attenuated by increasing the size of the array. Higher electric fields lead to intrinsic channeling, which is attributed to the disparate time scales for a rope-over-pulley collision and transverse diffusion between collisions. The onset of channeling is a gradual process, in agreement with extant Brownian dynamics simulation data. Even at weak electric fields, the electrophoretic mobility of λ-DNA in the array is considerably higher than would be expected if the DNA frequently collided with the posts.
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87.80.Kc Electrochemical techniques
82.45.Tv Bioelectrochemistry
87.14.gk DNA
87.15.Vv Diffusion

Dielectrophoretic separation of colorectal cancer cells

Fang Yang, Xiaoming Yang, Hong Jiang, Phillip Bulkhaults, Patricia Wood, William Hrushesky, and Guiren Wang

Biomicrofluidics 4, 013204 (2010); http://dx.doi.org/10.1063/1.3279786 (13 pages) | Cited 12 times

Online Publication Date: 12 January 2010

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Separation of colorectal cancer cells from other biological materials is important for stool-based diagnosis of colorectal cancer. In this paper, we use conventional dielectrophoresis in a microfluidic chip to manipulate and isolate HCT116 colorectal cancer cells. It is noticed that at a particular alternating current frequency band, the HCT116 cells are clearly deflected to a side channel from the main channel after the electric activation of an electrode pair. This motion caused by negative dielectrophoresis can be used to simply and rapidly separate cancer cells from other cells. In this manuscript, we report the chip design, flow conditions, dielectrophoretic spectrum of the cancer cells, and the enrichment factor of the colorectal cancer cells from other cells.
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87.85.Ox Biomedical instrumentation and transducers, including micro-electro-mechanical systems (MEMS)
85.85.+j Micro- and nano-electromechanical systems (MEMS/NEMS) and devices
87.19.xj Cancer
87.17.-d Cell processes

Designing a sensitive and quantifiable nanocolloid assay with dielectrophoretic crossover frequencies

Sagnik Basuray and Hsueh-Chia Chang

Biomicrofluidics 4, 013205 (2010); http://dx.doi.org/10.1063/1.3294575 (11 pages) | Cited 9 times

Online Publication Date: 22 January 2010

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Dielectrophoretic nanocolloid assay is a promising technique for sensitive molecular detection and identification, as target molecule hybridization onto the probe-functionalized nanocolloids can change their surface conductance and consequently their dielectrophoretic crossover frequencies. Thus, instead of relying on surface charge density increase after hybridization, as in many capacitive and field effect transistor impedance sensing techniques, the current assay utilizes the much larger surface conductance (and dielectrophoresis crossover frequency) changes to effect sensitive detection. Herein, we present a Poisson–Boltzmann theory for surfaces with finite-size molecular probes that include the surface probe conformation, their contribution to surface charge with a proper delineation of the slip and Stern planes. The theory shows that the most sensitive nanocolloid molecular sensor corresponds to a minimum in the dielectrophoretic crossover frequency with respect to the bulk concentration of the molecular probes (oligonucleotides in our case) during nanocolloid functionalization. This minimum yields the lowest number of functionalized probes that are also fully stretched because of surface probe-probe interaction. Our theory provides the surface-bulk oligonucleotide concentration isotherm and a folding number for the surface oligonucleotide conformation from the crossover frequency, the zeta potential, and the hydrodynamic radius data.
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87.80.-y Biophysical techniques (research methods)
85.85.+j Micro- and nano-electromechanical systems (MEMS/NEMS) and devices
87.14.gk DNA
87.15.-v Biomolecules: structure and physical properties
82.70.Dd Colloids

The current-voltage relation for electropores with conductivity gradients

Jianbo Li and Hao Lin

Biomicrofluidics 4, 013206 (2010); http://dx.doi.org/10.1063/1.3324847 (17 pages) | Cited 3 times

Online Publication Date: 1 March 2010

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In electroporation, an electric field transiently permeabilizes the cell membrane to gain access to the cytoplasm, and to deliver active agents such as DNA, proteins, and drug molecules. Past work suggests that the permeabilization is caused by the formation of aqueous, conducting pores on the lipid membrane, which are also known as electropores. The current-voltage relation across the membrane-bound pores is critical for understanding and predicting electroporation. In this work, we solve the Nernst–Planck equations in a geometry encompassing an isolated electropore to investigate this relation. In particular, we study cases where the intra- and extracellular electrical conductivities differ. We first derive an analytical solution, which is subsequently validated with a direct numerical simulation using a finite volume method. The main result of the current work is a formula for the effective pore resistance as a function of the pore radius, the membrane thickness, and the intra- and extracellular conductivities. This formula can be incorporated into whole-cell or planar-membrane electroporation models for system-level prediction and understanding.
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87.50.cj Electroporation/membrane effects
87.16.dp Transport, including channels, pores, and lateral diffusion
87.14.E- Proteins
87.14.gk DNA
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Mixing enhancement in microfluidic channel with a constriction under periodic electro-osmotic flow

Chun Yee Lim, Yee Cheong Lam, and Chun Yang

Biomicrofluidics 4, 014101 (2010); http://dx.doi.org/10.1063/1.3279790 (18 pages) | Cited 7 times

Online Publication Date: 7 January 2010

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We present a new approach to enhance mixing in T-type micromixers by introducing a constriction in the microchannel under periodic electro-osmotic flow. Two sinusoidal ac electric fields with 180° phase difference and similar dc bias are applied at the two inlets. The out of phase ac electric field induces oscillation of fluid interface at the junction of the two inlet channels and the constriction. Due to the constriction introduced at the junction, fluids from these two inlets form alternative plugs at the constricted channel. These plugs of fluids radiate downstream from the constriction into the large channel and form alternate thin crescent-shaped layers of fluids. These crescent-shaped layers of fluids increase tremendously the contact surface area between the two streams of fluid and thus enhance significantly the mixing efficiency. Experimental results and mixing mechanism analysis show that amplitude and frequency of the ac electric field and the length of the constriction govern the mixing efficiency.
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07.10.Cm Micromechanical devices and systems
47.65.-d Magnetohydrodynamics and electrohydrodynamics
66.10.C- Diffusion and thermal diffusion
47.61.Ne Micromixing
47.60.Dx Flows in ducts and channels
47.85.Np Fluidics

Dielectrophoretic spectra of translational velocity and critical frequency for a spheroid in traveling electric field

Sakshin Bunthawin, Pikul Wanichapichart, Adisorn Tuantranont, and Hans G. L. Coster

Biomicrofluidics 4, 014102 (2010); http://dx.doi.org/10.1063/1.3294082 (13 pages) | Cited 2 times

Online Publication Date: 13 January 2010

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An analysis has been made of the dielectrophoretic (DEP) forces acting on a spheroidal particle in a traveling alternating electric field. The traveling field can be generated by application of alternating current signals to an octapair electrode array arranged in phase quadrature sequence. The frequency dependent force can be resolved into two orthogonal forces that are determined by the real and the imaginary parts of the Clausius–Mossotti factor. The former is determined by the gradient in the electric field and directs the particle either toward or away from the tip of the electrodes in the electrode array. The force determined by the imaginary component is in a direction along the track of the octapair interdigitated electrode array. The DEP forces are related to the dielectric properties of the particle. Experiments were conducted to determine the DEP forces in such an electrode arrangement using yeast cells (Saccharomyces cervisiate TISTR 5088) with media of various conductivities. Experimental data are presented for both viable and nonviable cells. The dielectric properties so obtained were similar to those previously reported in literature using other DEP techniques.
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87.80.Ek Mechanical and micromechanical techniques
87.17.-d Cell processes
85.85.+j Micro- and nano-electromechanical systems (MEMS/NEMS) and devices
87.50.ch Electrophoresis/dielectrophoresis and other mechanical effects
82.45.-h Electrochemistry and electrophoresis

Rapid separation of bacteriorhodopsin using a laminar-flow extraction system in a microfluidic device

Yun Suk Huh, Chang-Moon Jeong, Ho Nam Chang, Sang Yup Lee, Won Hi Hong, and Tae Jung Park

Biomicrofluidics 4, 014103 (2010); http://dx.doi.org/10.1063/1.3298608 (10 pages) | Cited 1 time

Online Publication Date: 27 January 2010

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A protein separation technology using the microfluidic device was developed for the more rapid and effective analysis of target protein. This microfluidic separation system was carried out using the aqueous two-phase system (ATPS) and the ionic liquid two-phase system (ILTPS) for purification method of the protein sample, and the three-flow desalting system was used for the removal of salts from the sucrose-rich sample. Partitioning of the protein sample was observed in ATPS or ILTPS with the various pHs. The microdialysis system was applied to remove small molecules, such as sucrose and salts in the microfluidic channel with the different flow rates of buffer phase. A complex purification method, which combines microdialysis and ATPS or ILTPS, was carried out for the effective purification of bacteriorhodopsin (BR) from the purple membrane of Halobacterium salinarium, which was then analyzed by sodium dodecyl sulfatepolyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight. Furthermore, we were able to make a stable three-phase flow controlling the flow rate in the microfluidic channel. Our complex purification methods were successful in purifying and recovering the BR to its required value.
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87.80.Ek Mechanical and micromechanical techniques
87.14.E- Proteins
47.85.Np Fluidics
47.60.Dx Flows in ducts and channels
47.55.-t Multiphase and stratified flows
87.15.Tt Electrophoresis

Effect of electrical double layer on electric conductivity and pressure drop in a pressure-driven microchannel flow

Heng Ban, Bochuan Lin, and Zhuorui Song

Biomicrofluidics 4, 014104 (2010); http://dx.doi.org/10.1063/1.3328091 (13 pages) | Cited 2 times

Online Publication Date: 25 February 2010

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The effect of an electrical double layer (EDL) on microchannel flow has been studied widely, and a constant bulk electric conductivity is often used in calculations of flow rate or pressure drop. In our experimental study of pressure-driven micropipette flows, the pipette diameter is on the same order of magnitude as the Debye length. The overlapping EDL resulted in a much higher electric conductivity, lower streaming potential, and lower electroviscous effect. To elucidate the effect of overlapping EDL, this paper developed a simple model for water flow without salts or dissolved gases (such as CO2) inside a two-dimensional microchannel. The governing equations for the flow, the Poisson, and Nernst equations for the electric potential and ion concentrations and the charge continuity equation were solved. The effects of overlapping EDL on the electric conductivity, velocity distribution, and overall pressure drop in the microchannel were quantified. The results showed that the average electric conductivity of electrolyte inside the channel increased significantly as the EDL overlaps. With the modified mean electric conductivity, the pressure drop for the pressure-driven flow was smaller than that without the influence of the EDL on conductivity. The results of this study provide a physical explanation for the observed decrease in electroviscous effect for microchannels when the EDL layers from opposing walls overlap.
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87.85.Ng Biological signal processing
02.60.Lj Ordinary and partial differential equations; boundary value problems

Modeling of dielectrophoretic transport of myoglobin molecules in microchannels

Naga Siva Kumar Gunda and Sushanta Kumar Mitra

Biomicrofluidics 4, 014105 (2010); http://dx.doi.org/10.1063/1.3339773 (20 pages) | Cited 2 times

Online Publication Date: 1 March 2010

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Myoglobin is one of the premature identifying cardiac markers, whose concentration increases from 90 pg/ml or less to over 250 ng/ml in the blood serum of human beings after minor heart attack. Separation, detection, and quantification of myoglobin play a vital role in revealing the cardiac arrest in advance, which is the challenging part of ongoing research. In the present work, one of the electrokinetic approaches, i.e., dielectrophoresis (DEP), is chosen to separate the myoglobin. A mathematical model is developed for simulating dielectrophoretic behavior of a myoglobin molecule in a microchannel to provide a theoretical basis for the above application. This model is based on the introduction of a dielectrophoretic force and a dielectric myoglobin model. A dielectric myoglobin model is developed by approximating the shape of the myoglobin molecule as sphere, oblate, and prolate spheroids. A generalized theoretical expression for the dielectrophoretic force acting on respective shapes of the molecule is derived. The microchannel considered for analysis has an array of parallel rectangular electrodes at the bottom surface. The potential and electric field distributions are calculated using Green’s theorem method and finite element method. These results also compared to the Fourier series method, closed form solutions by Morgan et al. [J. Phys. D: Appl. Phys. 34, 1553 (2001) ] and Chang et al. [J. Phys. D: Appl. Phys. 36, 3073 (2003) ]. It is observed that both Green’s theorem based analytical solution and finite element based numerical solution for proposed model are closely matched for electric field and square electric field gradients. The crossover frequency is obtained as 40 MHz for given properties of myoglobin and for all approximated shapes of myoglobin molecule. The effect of conductivity of medium and myoglobin on the crossover frequency is also demonstrated. Further, the effect of hydration layer on the crossover frequency of myoglobin molecules is also presented. Both positive and negative DEP effects on myoglobin molecules are obtained by switching the frequency of applied electric field. The effect of different shapes of myoglobin on DEP force is studied and no significant effect on DEP force is observed. Finally, repulsion of myoglobin molecules from the electrode plane at 1 KHz frequency and 10 V applied voltage is observed. These results provide the ability of applying DEP force for manipulating nanosized biomolecules such as myoglobin.
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87.15.-v Biomolecules: structure and physical properties
87.14.E- Proteins
87.15.Pc Electronic and electrical properties
87.85.gf Fluid mechanics and rheology
87.19.U- Hemodynamics

Pressure driven spinning: A multifaceted approach for preparing nanoscaled functionalized fibers, scaffolds, and membranes with advanced materials

Suwan N. Jayasinghe and Nicolai Suter

Biomicrofluidics 4, 014106 (2010); http://dx.doi.org/10.1063/1.3328092 (13 pages) | Cited 3 times

Online Publication Date: 2 March 2010

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Electrospinning, a flexible jet-based fiber, scaffold, and membrane fabrication approach, has been elucidated as having significance to the heath sciences. Its capabilities have been most impressive as it possesses the ability to spin composite fibers ranging from the nanometer to the micrometer scale. Nonetheless, electrospinning has limitations and hazards, negating its wider exploration, for example, the inability to handle highly conducting suspensions, to its hazardous high voltage. Hence, to date electrospinning has undergone an exhaustive research regime to a point of cliché. Thus, in the work reported herein we unveil a competing technique to electrospinning, which has overcome the above limitations and hazards yet comparable in capabilities. The fiber preparation approach unearthed herein is referred to as “pressure driven spinning (PDS).” The driving mechanism exploited in this fiber spinning process is the pressurized by-pass flow. This mechanism allows the drawing of either micro- or nanosized fibers while processing polymeric suspensions containing a wide range of advanced materials spanning structural, functional, and biological entities. Similar to electrospinning if the collection time of these continuous formed fibers is varied, composite scaffolds and membranes are generated. In keeping with our interests, multicompositional structural entities such as these could have several applications in biology and medicine, for example, ranging from the development of three-dimensional cultures (including disease models) to the development of synthetic tissues and organ structures to advanced approaches for controlled and targeted therapeutics.
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87.16.D- Membranes, bilayers, and vesicles
81.20.-n Methods of materials synthesis and materials processing

Small volume laboratory on a chip measurements incorporating the quartz crystal microbalance to measure the viscosity-density product of room temperature ionic liquids

N. Doy, G. McHale, M. I. Newton, C. Hardacre, R. Ge, J. M. MacInnes, D. Kuvshinov, and R. W. Allen

Biomicrofluidics 4, 014107 (2010); http://dx.doi.org/10.1063/1.3353379 (7 pages) | Cited 2 times

Online Publication Date: 8 March 2010

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A microfluidic glass chip system incorporating a quartz crystal microbalance (QCM) to measure the square root of the viscosity-density product of room temperature ionic liquids (RTILs) is presented. The QCM covers a central recess on a glass chip, with a seal formed by tightly clamping from above outside the sensing region. The change in resonant frequency of the QCM allows for the determination of the square root viscosity-density product of RTILs to a limit of ∼ 10 kg m−2 s−0.5. This method has reduced the sample size needed for characterization from 1.5 ml to only 30 μl and allows the measurement to be made in an enclosed system.
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82.45.Gj Electrolytes
82.80.-d Chemical analysis and related physical methods of analysis
85.85.+j Micro- and nano-electromechanical systems (MEMS/NEMS) and devices
85.50.-n Dielectric, ferroelectric, and piezoelectric devices
66.20.-d Viscosity of liquids; diffusive momentum transport

Band-broadening suppressed effect in long turned geometry channel and high-sensitive analysis of DNA sample by using floating electrokinetic supercharging on a microchip

Zhongqi Xu, Kenji Murata, Akihiro Arai, and Takeshi Hirokawa

Biomicrofluidics 4, 014108 (2010); http://dx.doi.org/10.1063/1.3366719 (12 pages) | Cited 1 time

Online Publication Date: 12 March 2010

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A featured microchip owning three big reservoirs and long turned geometry channel was designed to improve the detection limit of DNA fragments by using floating electrokinetic supercharging (FEKS) method. The novel design matches the FEKS preconcentration needs of a large sample volume introduction with electrokinetic injection (EKI), as well as long duration of isotachophoresis (ITP) process to enrich low concentration sample. In the curved channel [ ∼ 45.6 mm long between port 1 (P1) and the intersection point of two channels], EKI and ITP were performed while the side port 3 (P3) was electrically floated. The turn-induced band broadening with or without ITP process was investigated by a computer simulation (using CFD-ACE+ software) when the analytes traveling through the U-shaped geometry. It was found that the channel curvature determined the extent of band broadening, however, which could be effectively eliminated by the way of ITP. After the ITP-stacked zones passed the intersection point from P1, they were rapidly destacked for separation and detection from ITP to zone electrophoresis by using leading ions from P3. The FEKS carried on the novel chip successfully contributed to higher sensitivities of DNA fragments in comparison with our previous results realized on either a single channel or a cross microchip. The analysis of low concentration 50 bp DNA step ladders (0.23 μg/ml after 1500-fold diluted) was achieved with normal UV detection at 260 nm. The obtained limit of detections (LODs) were on average 100 times better than using conventional pinched injection, down to several ng/ml for individual DNA fragment.
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87.15.Tt Electrophoresis
87.14.gk DNA

Simultaneous measurement of concentrations and velocities of submicron species using multicolor imaging and microparticle image velocimetry

Jing-Tang Yang, Yu-Hsuan Lai, Wei-Feng Fang, and Miao-Hsing Hsu

Biomicrofluidics 4, 014109 (2010); http://dx.doi.org/10.1063/1.3366721 (11 pages) | Cited 2 times

Online Publication Date: 15 March 2010

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We propose a novel approach to resolve simultaneously the distributions of velocities and concentration of multiple, submicron species in microfluidic devices using microparticle image velocimetry, and particle counting. Both two-dimensional measurement and three-dimensional analysis of flow fields, from the stacked images, are achieved on applying a confocal fluorescence microscope. The displacements of all seeding particles are monitored to determine the overall velocity field, whereas the multicolor particles are counted and analyzed individually for each color to reveal the distributions of concentration and velocity of each species. A particle-counting algorithm is developed to determine quantitatively the spatially resolved concentration. This simultaneous measurement is performed on a typical T-shaped channel to investigate the mixing of fluids. The results are verified with numerical simulation; satisfactory agreement is achieved. This measurement technique possesses reliability appropriate for a powerful tool to analyze multispecies mixing flows, two-phase flows, and biofluids in microfluidic devices.
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87.64.M- Optical microscopy
02.60.-x Numerical approximation and analysis
87.15.mq Luminescence

Induced charge electro-osmotic concentration gradient generator

Mranal Jain, Anthony Yeung, and K. Nandakumar

Biomicrofluidics 4, 014110 (2010); http://dx.doi.org/10.1063/1.3368991 (13 pages)

Online Publication Date: 23 March 2010

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Biomolecule gradients play an important role in the understanding of various biological processes. Typically, biological cells are exposed to linear and nonlinear concentration gradients and their response is studied for understanding cell growth, cell migration, and cell differentiation mechanisms. Recent studies have demonstrated the use of microfluidic devices for precise and stable concentration gradient generation. However, most of the reported devices are geometrically complex and lack dynamic controllability. In this work, a novel microfluidic gradient generator is presented which utilizes the induced charge electro-osmosis (ICEO) by introducing conducting obstacle in the microchannel. With the ICEO flow component, significant transverse convection can be generated within the microchannel, which can, in turn, be used to create nonlinear as well as asymmetric gradients. The characteristics of the developed concentration gradient are dependent on the interplay between fixed charge electro-osmotic and ICEO flows. It is shown that the proposed device can switch between linear and nonlinear gradients by just altering the applied electric field. Finally, the formation of user-defined concentration profiles (linear, convex, and concave) is demonstrated by varying the conducting obstacle size.
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85.85.+j Micro- and nano-electromechanical systems (MEMS/NEMS) and devices
87.15.hj Transport dynamics
87.80.Ek Mechanical and micromechanical techniques
47.85.Np Fluidics
82.39.Wj Ion exchange, dialysis, osmosis, electro-osmosis, membrane processes
87.17.-d Cell processes
82.80.-d Chemical analysis and related physical methods of analysis

Cell death along single microfluidic channel after freeze-thaw treatments

Yuhui Li, Fen Wang, and Hao Wang

Biomicrofluidics 4, 014111 (2010); http://dx.doi.org/10.1063/1.3324869 (10 pages) | Cited 2 times

Online Publication Date: 25 March 2010

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Show Abstract
Cryotherapy is a prospective green method for malignant tumor treatment. At low temperature, the cell viability relates with the cooling rate, temperature threshold, freezing interface, as well as ice formation. In clinical applications, the growth of ice ball must reach a suitable size as cells could not be all killed at the ice periphery. The cell death ratio at the ice periphery is important for the control of the freezing destruction. The mechanisms of cryoinjury around the ice periphery need thorough understanding. In this paper, a primary freeze-thaw control was carried out in a cell culture microchip. A series of directional freezing processes and cell responses was tested and discussed. The temperature in the microchip was manipulated by a thermoelectric cooler. The necrotic and apoptotic cells under different cryotreatment (duration of the freezing process, freeze-thaw cycle, postculture, etc.) were stained and distinguished by propidium iodide and fluorescein isothiocyanate (FITC)-Annexin V. The location of the ice front was recorded and a cell death boundary which was different from the ice front was observed. By controlling the cooling process in a microfluidic channel, it is possible to recreate a sketch of biological effect during the process of simulated cryosurgery.
Show PACS
87.19.Pp Biothermics and thermal processes in biology
87.85.-d Biomedical engineering
47.85.Np Fluidics
87.17.-d Cell processes
07.20.Mc Cryogenics; refrigerators, low-temperature detectors, and other low-temperature equipment
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