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Droplet microfluidics (DMF) is a fluidic handling technology that enables precision control over dispensing and subsequent manipulation of droplets in the volume range of microliters to picoliters, on a micro-fabricated device. There are several different droplet actuation methods, all of which can generate external stimuli, to either actively or passively control the shape and positioning of fluidic droplets over patterned substrates. In this review article, we focus on the operation and utility of electro-actuation-based DMF devices, which utilize one or more micro-/nano-patterned substrates to facilitate electric field-based handling of chemical and/or biological samples. The underlying theory of DMF actuations, device fabrication methods and integration of optical and opto-electronic detectors is discussed in this review. Example applications of such electro-actuation-based DMF devices have also been included, illustrating the various actuation methods and their utility in conducting chip-based laboratory and clinical diagnostic assays.

Liquid dielectrophoresis (L-DEP), when deployed at microscopic scales on top of hydrophobic surfaces, offers novel ways of rapid and automated manipulation of very small amounts of polar aqueous samples for microfluidic applications and development of laboratory-on-a-chip devices. In this article we highlight some of the more recent developments and applications of L-DEP in handling and processing of various types of aqueous samples and reagents of biological relevance including emulsions using such microchip based surface microfluidic (SMF) devices. We highlighted the utility of these devices for on-chip bioassays including nucleic acid analysis. Furthermore, the parallel sample processing capabilities of these SMF devices together with suitable on- or off-chip detection capabilities suggest numerous applications and utility in conducting automated multiplexed assays, a capability much sought after in the high throughput diagnostic and screening assays.

A miniaturized microfluidic device was developed to facilitate electromanipulation of bacterial respiratory pathogens. The device comprises a microchip with circular aluminum electrodes patterned on glass, which is housed in a microfluidic system fabricated utilizing polydimethylsiloxane. The device provides sample preparation capability by exploiting positive dielectrophoresis (DEP) in conjunction with pulsed voltage for manipulation and disruption of Bordetella pertussis bacterial cells. Positive DEP capture of B. pertussis was successfully demonstrated utilizing 10 Vrms and 1 MHz ac fields. Application of dc pulses (300 V amplitude and 50 micros pulsewidth applied 1 s apart) across the aluminum electrodes resulted in electrodisruption and lysis of B. pertussis bacterial cells. Real-time polymerase chain reaction, a 2(3) factorial experimental design and transmission electron microscopy were used to evaluate bacterial cell manipulation and factors affecting bacterial cell disruption. The main factors affecting bacterial cell disruption were electric field strength, the electrical conductivity of the cell suspension sample, and the combined effect of number of pulses and sample conductivity. The bacterial deoxyribonucleic acid target remained undamaged as a result of DEP and cell lysis experimentation. Our findings suggest that a simple miniaturized microfluidic device can achieve important steps in sample preparation on-chip involving respiratory bacterial pathogens.

Twenty-four-hour ambulatory pH monitoring is an essential tool for diagnosing gastroesophageal reflux disease (GERD). Simultaneous impedance and pH monitoring of the esophagus improves the detection and characterization of GERD. Conventional catheter-based monitoring systems are uncomfortable and interfere with the normal activity of the patient. To overcome these disadvantages, different wireless esophageal monitoring systems have been proposed. A capsule containing sensors for impedance and pH monitoring with wireless communication capabilities is presented. A low cost miniature microcontroller was utilized for interfacing between the sensors and a wireless transmitter. The microcontroller program allowed efficient management of the electric power provided by a 3-V battery. Magnetic holding is proposed as an alternative to surgical affixation of the monitoring capsule. Permanent neodymium magnets separated by 27 cm successfully held the capsule in a test tube. Experimental results demonstrated that friction force can aid magnetic holding to overcome peristalsis. The proposed design efficiently detected acid and nonacid reflux. More research regarding the holding method and capsule packaging are necessary to optimize the mechanical performance of the proposed design in order to facilitate clinical testing on human subjects.

Strong frequency-dependent fluid flow has been observed near the surface of microelectrode arrays. Modeling this phenomenon has proven to be difficult, with existing theories unable to account for the qualitative trend observed in the frequency spectra of this flow. Using recent electrode polarization results, a more comprehensive model of the double layer on the electrode surface is used to obtain good theoretical agreement with experimental data. Copyright 2001 Academic Press.

The expressions obtained in the previous paper for electrode polarization are applied to a homogeneous planar electrode and a planar array of electrodes used in the generation of nonuniform fields. The effective far field experienced outside the double layer is computed for both electrodes, and sample spectra are provided. The effective far field expression contains the electrode impedance and the effects of concentration polarization due to the static double layer on the electrode generated by the zeta potential. The effective far field results are compact and contain simple integrals that can be evaluated numerically. Copyright 2000 Academic Press.

A novel dual-frequency excitation technique is introduced which permits investigation of the low-frequency dispersion of Canola plant protoplasts using feedback-controlled dielectrophoretic levitation. The upper and intermediate frequency spectra obtained using the new technique are generally consistent with previous work. However, below some cross-over frequency f(OL), the protoplasts exhibit an apparent positive dielectrophoretic response that is not predicted by conventional theory. This cross-over frequency is linearly related to suspension conductivity, virtually independent of the suspension pH, and inversely proportional to the square of the cell radius. Examination of the complex Clausius-Mossotti polarization coefficient reveals that the observed positive dielectrophoretic response can not be accounted for in terms of Maxwell-Wagner polarization associated with a conventional layered model for the protoplast. The failure of straightforward enhancements to the protoplast model in explaining the low frequency behavior may indicate the presence of an electrophoretic contribution to the net observable force on the particle. To account for such fluid mechanical effects, it will be necessary to modify the existing dielectrophoretic force formulation.

The contributions of pulse-induced dipole-dipole interaction to the total pressure acting normal to the membranes of closely positioned pronase treated human erythrocytes during electrofusion was calculated. The total pressure was modeled as the sum of pressures arising from membrane potential and dipole-dipole attraction opposed by interbilayer repulsion. The dipole-dipole interaction was derived from the experimentally obtained cell polarizability. The threshold electric field amplitude necessary for fusion of pronase-treated human erythrocytes was experimentally obtained at various combinations of pulse duration, frequency, and the conductivity of the external medium. The theoretical values of the critical electric field amplitude compared favorably to the experimentally obtained threshold field amplitudes. Fusion by dc pulses may be primarily attributed to attainment of sufficiently high membrane potentials. However, with decreasing external conductivity and increasing sinusoidal pulse frequency (100 kHz-2.5 MHz), the induced dipole-dipole interactions provide the principal driving force for membrane failure leading to fusion.

In this paper we have utilized the principle of dielectrophoresis (DEP) to develop an apparatus to stably levitate single biological cells using a digital feedback control scheme. Using this apparatus, the positive DEP spectra of both Canola plant protoplast and ligament fibroblast cells have been measured over a wide range of frequencies (1 kHz to 50 MHz) and suspending medium conductivities (11-800 muS/cm). The experimental data thus obtained have been interpreted in terms of a simple spherical cell model. Furthermore, utilizing such a model, we have shown that various cellular parameters of interest can be readily obtained from the measured DEP levitation spectrum. Specifically, the effective membrane capacitance of single cells has been determined. Values of 0.47 +/- 0.03 muF/cm2 for Canola protoplasts and 1.52 +/- 0.26 muF/cm2 for ligament fibroblasts thus obtained are consistent with those determined by other existing electrical methods.

Droplet microfluidics has in recent years found a wide range of analytical and bioanalytical applications. In droplet microfluidics, the samples that are discretized into droplets within the devices are predominantly loaded through tubings, but such tubing-based sample loading has drawbacks such as limited scalability for processing many samples, difficulty for automation, and sample wastage. While advances in autosamplers have alleviated some of these drawbacks, sample loading that can instead obviate tubings offers a potentially promising alternative but has been underexplored. To fill the gap, we introduce herein a droplet device that features a new Tubing Eliminated Sample Loading Interface (TESLI). TESLI integrates a network of programmable pneumatic microvalves that regulate vacuum and pressure sources so that successive sub-microliter samples can be directly spotted onto the open-to-atmosphere TESLI inlet, vacuumed into the device, and pressurized into nanoliter droplets within the device with minimal wastage. The same vacuum and pressure regulation also endows TESLI with cleaning and sample switching capabilities, thus enabling scalable processing of many samples in succession. Moreover, we implement a pair of TESLIs in our device to parallelize and alternate their operation as means to minimizing idle time. For demonstration, we use our device to successively process 44 samples into droplets—a number that can further scale. Our results demonstrate the feasibility of tubing-free sample loading and a promising approach for advancing droplet microfluidics. [ABSTRACT FROM AUTHOR]

Label-free and gentle separation of cell stages with desired target properties from mixed stage populations are a major research task in modern biotechnological cultivation process and optimization of micro algae. The reported microfluidic sorter system (MSS) allows the subsequent investigation of separated subpopulations. The implementation of a viability preserving MSS is shown for separation of late stage 1 Haematococcus pluvialis (HP) cells form a mixed stage population. The MSS combines a three-step flow focusing unit for aligning the cells in single file transportation mode at the center of the microfluidic channel with a pure hydrodynamic sorter structure for cell sorting. Lateral displacement of the cells into one of the two outlet channels is generated by piezo-actuated pump chambers. In-line decision making for sorting is based on a user-definable set of image features and properties. The reported MSS significantly increased the purity of target cells in the sorted population (94%) in comparison to the initial mixed stage population (19%). [ABSTRACT FROM AUTHOR]

Isomotive dielectrophoresis (isoDEP) is a unique electric field such that the gradient of the field-squared (∇ E rms 2 ) is constant, resulting a uniform dielectrophoretic force. The current status of isoDEP is presented in this review, and we will highlight the progress that has been achieved over the past 60 years in various avenues of isoDEP since H.A. Pohl initially described its premise. This article will discuss its applications and describe the various configurations of generating an isomotive force. Since H.A. Pohl introduced the theory of isoDEP, numerous authors have implemented isoDEP as a tool for the manipulation, sorting, separation, and characterization of polarizable particles without the need for biochemical labels or other bioengineered tagging. The growing field of microfluidics and electrokinetics has renewed interest in isoDEP, particularly for analytical characterization or separation of particles. Recent work has demonstrated that isoDEP can address some unmet needs for biomedical applications including single-cell analysis; moreover, advances in throughput as well as combining characterization and separation simultaneously will add significant value to isoDEP. [ABSTRACT FROM AUTHOR]

The multi-disciplinary field of microfluidics has the potential to provide solutions to a diverse set of problems. It offers the advantages of high-throughput, continuous, rapid and expeditious analysis requiring minute quantities of sample. However, even as this field has yielded many mass-manufacturable and cost-efficient point-of-care devices, its direct and practical applications into the field of disease diagnostics still remain limited and largely overlooked by the industry. This review focuses on the phenomenon of hydrodynamic focusing and its potential to materialize solutions for appropriate diagnosis and prognosis. The study aims to look beyond its intended cytometric applications and focus on unambiguous disease detection, monitoring, drug delivery, studies conducted on DNA and highlight the instances in the scientific literature that have proposed such approach. [ABSTRACT FROM AUTHOR]

This review aims at focusing on the use of microneedles (MNs) as an emerging novel drug delivery carrier for an effective treatment in diabetic patients. There are many limitations in various modes of delivery such as oral, subcutaneous, nasal, and other modes which cause pain and have many other side effects. Hence, this drug delivery research has found to have tremendous potential in combining both the diagnostic and therapeutic elements, thus treating diabetes in a better way. Most glucose-sensing techniques and conventional insulin therapies are engaged in the transfer of physical entities through the skin. MN- based drug delivery system can accomplish in an noninvasive or minimally invasive manner which can be an add on advantage towards pain-free administration, easy handling, discrete, continuous as well as providing a controlled release system. Hence, the review addresses on the current advancement of this bioengineered system like MNs, constituting a "smart" system specifically for autonomous diabetes therapy. [ABSTRACT FROM AUTHOR]

Personalized medicine - healthcare based on individual genetic variation - has the potential to transform the way healthcare is delivered to patients. The promise of personalized medicine has been predicated on the predictive and diagnostic power of genomic and proteomic biomarkers. Biomarker screening may help improve health outcomes, for example, by identifying individuals' susceptibility to diseases and predicting how patients will respond to drugs. Microfluidic droplet technology offers an exciting opportunity to revolutionize the accessibility of personalized medicine. A framework for the role of droplet microfluidics in biomarker detection can be based on two main themes. Emulsion-based microdroplet platforms can provide new ways to measure and detect biomolecules. In addition, microdroplet platforms facilitate high-throughput screening of biomarkers. Meanwhile, surface-based droplet platforms provide an opportunity to develop miniaturized diagnostic systems. These platforms may function as portable benchtop environments that dramatically shorten the transition of a benchtop assay into a point-of-care format. [ABSTRACT FROM AUTHOR]

By simultaneously subjecting microbial cells to high amplitude pulsed electric fields and flash heating of the cell suspension fluid, effective release of intracellular contents was achieved. The synergistic effect of the applied electric field and elevated temperature on cell lysis in a flow-through device was demonstrated for Gram-negative and Gram-positive bacteria, and Mycobacterium species. The resulting lysate is suitable for downstream nucleic acid amplification and detection without requiring further preparation. The lysis chamber employs surface enhanced blocking electrodes which possess an etched micro-structured surface and a thin layer of dielectric metal oxide which provides a large effective area and blocks transmission of electrical current. The surface enhanced blocking electrodes enable simultaneous suppression of the rapid onset of electric field screening in the bulk of the cell suspension medium and avoidance of undesired electrochemical processes at the electrode-electrolyte interface. In addition the blocking layer ensures the robustness of the cell lysis device in applications involving prolonged flow-through processing of the microbial cells. [ABSTRACT FROM AUTHOR]

Background: Microelectrode dieletrophoresis capture of live cells has been explored in animal and cellular models ex-vivo. Currently, there is no clinical data available regarding the safety and efficacy of dielectrophoresis (DEP) buffers and microcurrent manipulation in humans, despite copious pre-clinical studies suggesting its safety. The purpose of this study was to determine if DEP isolation of SVF using minimal manipulation methods is safe and efficacious for use in humans using the hand lipotransfer model. Methods: Autologous stromal vascular fraction cells (SVF) were obtained from lipoaspirate by collagenase digestion and centrifugation. The final mixture of live and dead cells was further processed using a custom DEP microelectrode array and microcurrent generator to isolate only live nucleated cells. Lipotransfer was completed using fat graft enhanced with either standard processed SVF (control) versus DEP filtered SVF (experimental). Spectral photography, ultrasound and biometric measurements were obtained at post operatively days 1, 4, 7, 14, 30, 60 and 90. Results: The DEP filter was capable of increasing SVF viability counts from 74.3 ± 2.0% to 94.7 ± 2.1%. Surrogate markers of inflammation (temperature, soft tissue swelling, pain and diminished range of motion) were more profound on the control hand. Clinical improvement in hand appearance was appreciated in both hands, though the control hand exclusively sustained late phase erosive skin breaks on post operative day 7. No skin breaks were appreciated on the DEP-SVF treated hand. Early fat engraftment failure was noted on the control hand thenar web space at 3 months post surgery. Discussion: No immediate hypersensitivity or adverse reaction was appreciated with the DEP-SVF treated hand. In fact, the control hand experienced skin disruption and mild superficial cellulitis, whereas the experimental hand did not experience this complication, suggesting a possible "protective" effect with DEP filtered SVF. Late ultrasound survey revealed larger and more frequent formation of oil cysts in the control hand, also suggesting greater risk of engraftment failure with standard lipotransfer. Conclusion: Clinical DEP appears safe and efficacious for human use. The DEP microelectrode array was found to be versatile and robust in efficiently isolating live SVF cells from dead cells and cellular debris in a time sensitive clinical setting. [ABSTRACT FROM AUTHOR]

Background: Custom synthesized DNA is in high demand for synthetic biology applications. However, current technologies to produce these sequences using assembly from DNA oligonucleotides are costly and labor-intensive. The automation and reduced sample volumes afforded by microfluidic technologies could significantly decrease materials and labor costs associated with DNA synthesis. The purpose of this study was to develop a gene assembly protocol utilizing a digital microfluidic device. Toward this goal, we adapted bench-scale oligonucleotide assembly methods followed by enzymatic error correction to the Mondrian™ digital microfluidic platform. Results: We optimized Gibson assembly, polymerase chain reaction (PCR), and enzymatic error correction reactions in a single protocol to assemble 12 oligonucleotides into a 339-bp double- stranded DNA sequence encoding part of the human influenza virus hemagglutinin (HA) gene. The reactions were scaled down to 0.6-1.2 μL. Initial microfluidic assembly methods were successful and had an error frequency of approximately 4 errors/kb with errors originating from the original oligonucleotide synthesis. Relative to conventional benchtop procedures, PCR optimization required additional amounts of MgCl2, Phusion polymerase, and PEG 8000 to achieve amplification of the assembly and error correction products. After one round of error correction, error frequency was reduced to an average of 1.8 errors kb− 1. Conclusion: We demonstrated that DNA assembly from oligonucleotides and error correction could be completely automated on a digital microfluidic (DMF) platform. The results demonstrate that enzymatic reactions in droplets show a strong dependence on surface interactions, and successful on-chip implementation required supplementation with surfactants, molecular crowding agents, and an excess of enzyme. Enzymatic error correction of assembled fragments improved sequence fidelity by 2-fold, which was a significant improvement but somewhat lower than expected compared to bench-top assays, suggesting an additional capacity for optimization. [ABSTRACT FROM AUTHOR]