Thesis (M.S.)--University of Akron, Dept. of Mechanical Engineering, 2008. / "December, 2008." Title from electronic thesis title page (viewed 12/9/2009) Advisor, Jiang John Zhe; Faculty Readers, Joan Carletta, Dane Quinn; Department Chair, Celal Batur; Dean of the College, George K. Haritos; Dean of the Graduate School, George R. Newkome. Includes bibliographical references.

As digital microfluidics has continued to mature since its advent in the early 1980's, an increase in new and novel applications of this technology have been developed. However, even as this technology has become more common place, a consensus on the physics and force models of the motion of the contact line between the fluid, substrate, and ambient has not been reached. This uncertainty along with the dependence of the droplet geometry on the force to cause its motion has directed much of the research at specific geometries and droplet actuation methods. The goal of this thesis is to help characterize the components of the friction force which opposes droplet motion as a one dimensional system model based upon simple system parameters independent from the actuation method. To this end, the force opposing the motion of a droplet under a thin rectangular glass cover slip was measured for varying cover slip dimensions (widths, length), gap height between the cover slip and substrate, and bulk droplet velocity. The stiffness of the droplet before droplet motion began, the force at which the motion initiated, and the steady-state force opposing the droplet motion were measured. The data was then correlated to hypothesized equations and compared to simple models accounting for the forces due to the contact angle hysteresis, contact line friction, and viscous losses. It was found that the stiffness, breakaway force, and steady-state force of the droplet could be correlated to with an error standard deviation of 8 %, 14%, and 10 % respectively. Much of the error was due to an unexpected height dependence for the breakaway and steady-state forces and testing error associated with the velocity. The models for the stiffness and breakaway force over predicted the results by 36% and 16% respectively. During testing, viii stability issues with the cover slip were observed and simple dye testing was conducted to visualize the droplet flow field.

In recent years, BioMEMS/NEMS have been primary elements associated with the research and development efforts in the bioengineering area. International and federal funding has effected an enormous increase in the development of state-of-the-art bioengineering and biomedical technologies. Most of the BioMEMS/NEMS related applications are associated with diagnostics, sensing and detection. Procedures for separation and manipulation of biological components play a paramount role in the function of these bioengineering mechanisms. This research was concerned with the development of a novel BioMEMS device for cell manipulation. The functioning of the device is based on the use of thermally responsive polymer networks, which differs dramatically from existing approaches. This approach is cost effective, requires low power and uses a minimal amount of on-device area, which makes it suitable for personal medical diagnostics and battle field scenarios. The device integrates the technologies associated with reversibly binding surfaces and dielectrophoresis, (DEP). The DEP field drives a sample into contact with a binding surface. This surface can be controlled to provide different levels of target selectivity. This system provides a separation strategy that does not suffer from fouling issues. The binding surfaces are fabricated from LCST polymers. The LCST polymers experience hydration-dehydration changes in response to temperature fluctuations. Therefore, separation efficiency can be "dialed in" as a function of temperature to prompt the selection of targets. Furthermore, size-exclusion "trenches" were patterned into the binding surfaces. The trenches permit the passage of the small objects in order to provide size-exclusion separations. In order to expand the discrimination size range from the micron to the submicron scale, two techniques for submicron patterning of cross-linked reversibly binding surfaces were investigated. The patterning techniques associated with electron-beam lithography and the combination of softlithography and a focused ion beam patterning were found to generate well-defined patterns that retained their thermo-responsiveness. The combination of DEP and reversibly binding surfaces for bio-particle manipulation is a significant contribution to microfluidic based separations in BioMEMS/NEMS. The developments associated with this research provide a novel technology platform that facilitates separations, which would be difficult to achieve by any other existing methods.

When operating microsystems in harsh environments, many conventionally used techniques are limiting. Further, depending on if the demands arise from the environment or the conditions inside the system, different approaches have to be used. This thesis deals with the challenges encountered when microsystems are used at high pressures and high temperatures. For microsystems operating at harsh conditions, many parameters will vary extensively with both temperature and pressure, and to maintain control, these variations needs to be well understood. Covered within this thesis is the to-date strongest membrane micropump, demonstrated to pump against back-pressures up to 13 MPa, and a gas-tight high pressure valve that manages pressures beyond 20 MPa. With the ability to manipulate fluids at high pressures in microsystems at elevated temperatures, opportunities are created to use green solvents like supercritical fluids like CO2. To allow for a reliable and predictable operation in systems using more than one fluid, the behavior of the multiphase flow needs to be controlled. Therefore, the effect of varying temperature and pressure, as well as flow conditions were investigated for multiphase flows of CO2 and H2O around and above the critical point of CO2. Also, the influence of channel surface and geometry was investigated. Although supercritical CO2 only requires moderate temperatures, other supercritical fluids or reactions require much higher temperatures. The study how increasing temperature affects a system, a high-temperature testbed inside an electron microscope was created. One of the challenges for high-temperature systems is the interface towards room temperature components. To circumvent the need of wires, high temperature wireless systems were studied together with a wireless pressure sensing system operating at temperatures up to 1,000 °C for pressures up to 0.3 MPa. To further extend the capabilities of microsystems and combine high temperatures and high pressures, it is necessary to consider that the requirements differs fundamentally. Therefore, combining high pressures and high temperatures in microsystems results in great challenges, which requires trade-offs and compromises. Here, steel and HTCC based microsystems may prove interesting alternatives for future high performance microsystems.

The phenomenon of directed cell motion in response to external directional cues has drawn significant interest for more than a century, with the first recorded observations of bacterial chemotaxis at the end of the 19th century. Furthermore, movies generated by David Rogers while at Vanderbilt University of a peripheral blood neutrophil tracking a bacterium are a staple of any college biology class to demonstrate the phenomenon of eukaryotic chemotaxis. In just the last decade, our understanding of the biochemical mechanisms underlying the process of directed eukaryotic cell migration. As a result, several generalized processes have been identified, connecting multiple phenomena from cancer metastasis to axon guidance. Making further sense of the complex biochemical pathways requires both quantitative mathematical models and fine control over the external cellular environment. To this end, microfluidics has proven extremely useful, allowing for precise quantification of both the external environment and the cellular response.

Molecular diagnostics offer quick access to information for healthcare decision-making towards personalized therapeutics, but complicated procedures requiring extensive labor and infrastructure restrict their use. Droplet-based technologies can expand the accessibility of molecular diagnostics by miniaturizing devices, shortening sample-to-answer times, decreasing costs and increasing throughput. Methods for droplet manipulation are central to the automation of molecular diagnostics protocols. The innovative method, wire-guided droplet manipulation (WDM), is the actuation of liquid droplets in a hydrophobic milieu with a wire, or needle, guide. In this work, WDM is demonstrated for the automation of the polymerase chain reaction (PCR) on reprogrammable platforms for the diagnosis of cardiovascular infections. WDM is used to minimize thermal resistance by convective heat transfer for PCR amplification at a maximum speed of 8.67 s/cycle. The oil-water interfacial boundary is shown to passively partition molecular contaminants from sample matrices, including blood and heart valve tissue. Molecular self-assembly at the oil-water interface is used to increase PCR efficiency with blood in situ and is used as an innovative sensing modality for real-time monitoring of PCR amplification. Temperature feedback controlled droplet actuation is achieved by using a thermocouple loop as a functionalized wire-guide. Our novel methodology for real-time PCR, droplet-on-thermocouple silhouette real-time PCR (DOTS qPCR), utilizes interfacial effects to achieve droplet actuation, relief from PCR inhibitors and amplification sensing, for a sample-to-answer time as short as 3 min 30 s. DOTS qPCR addresses three major issues for rapid PCR—sample preparation, rapid thermocycling and sensitive real-time detection—on an inexpensive, disposable device with smartphone-based detection. In contrast, commercially available real-time PCR systems rely on fluorescence detection, have substantially higher threshold cycles, and require expensive optical components and extensive sample preparation. Due to the advantages of low threshold cycle detection we anticipate extending this technology towards trending biological research applications such as single cell, single nucleus, and single DNA molecule analyses, especially in droplet microfluidic platforms.

The use of smartphone for the detection of biological constituents is becoming a useful tool as a point-of-care (POC) device and diagnostics. When combined with microfluidic paper analytic devices (μPAD) and particle immunoassays, we have the ability to detect bacterial pathogens with sensitivity and specificity. Environmental conditions as well as variability in smartphone imaging and the cellulose in paper microfluidics however can sometimes easily interfere with the detection of small signal changes. Combining this issue with the detection of pathogens in blood (our model biological sample of interest) becomes difficult with such a platform because of the complexity of the sample matrix. However, in this research we take a novel approach at utilizing polystyrene’s auto-fluorescence and the high energy of UV LEDs in a particle immunoassay in order to increase our signal change. We first characterized how the smartphone actually responds to UV light (275-385 nm) with respect to the RGB components in its images. We were then able to determine a favorable response using the 385 nm UV LED. The detection of green fluorescence by polystyrene particles was possible by analyzing the smartphone’s image in the green channel. There was a significant difference in signal change with blood samples including polystyrene versus just blood samples with a normalized signal intensity change of 2.5 (150%). The detection of polystyrene fluorescence was translated into a field deployable prototype, where preliminary trials showed promising results in detecting Escherichia coli in blood with a current limit of detection of 50 CFU/ml. With further experimentation and optimization the limit of detection could be improved to 10 CFU/mL, making it a very useful tool in the detection of blood borne pathogens to prevent complications with onset bacteremia and the more serious cases of sepsis. This assay platform could provide an easy to use solution with detection in a short time (assay time of 1 min) compared to the lengthy blood culture monitoring or biomarker detection.

Particle-laden flow in a microchannel resulting in aggregation of microparticles was investigated to determine the dependence of the cluster growth rate on the following parameters: suspension void fraction, shear strain rate, and channel-height to particle-diameter ratio. The growth rate of an average cluster was found to increase linearly with suspension void fraction, and to obey a power-law relationships with shear strain rate as S^0.9 and channel-height to particle-diameter ratio as (h/d)^-3.5. Ceramic liposomal nanoparticles and silica microparticles were functionalized with antibodies that act as targeting ligands. The bio-functionality and physical integrity of the cerasomes were characterized. Surface functionalization allows cerasomes to deliver drugs with selectivity and specificity that is not possible using standard liposomes. The functionalized particle-target cell binding process was characterized using BT-20 breast cancer cells. Two microfluidic systems were used; one with both species in suspension, the other with cells immobilized inside a microchannel and particle suspension as the mobile phase. Effects of incubation time, particle concentration, and shear strain rate on particle-cell binding were investigated. With both species in suspension, the particle-cell binding process was found to be reasonably well-described by a first-order model. Particle desorption and cellular loss of binding affinity in time were found to be negligible; cell-particle-cell interaction was identified as the limiting mechanism in particle-cell binding. Findings suggest that separation of a bound particle from a cell may be detrimental to cellular binding affinity. Cell-particle-cell interactions were prevented by immobilizing cells inside a microchannel. The initial stage of particle-cell binding was investigated and was again found to be reasonably well-described by a first-order model. For both systems, the time constant was found to be inversely proportional to particle concentration. The second system revealed the time constant to obey a power-law relationship with shear strain rate as τ∝S^.37±.06. Under appropriate scaling, the behavior displayed in both systems is well-described by the same exponential curve. Identification of the appropriate scaling parameters allows for extrapolation and requires only two empirical values. This could provide a major head-start in any dosage optimization studies.

Electrophoresis separation corresponds to the motion and separation of dispersed particles under the influence of a constant electric field. In molecular biology, electrophoresis separation plays a major role in identifying, quantifying and studying different biological samples such as proteins, peptides, RNA acids, and DNA. In electrophoresis separation, different characteristics of particles, such as charge to mass ratio, size, and pI, can be used to separate and isolate those particles. For very complex samples, two or more characteristics can be combined to form a multi-dimensional electrophoresis separation system, significantly improving separation efficiency. Much effort has been devoted in recent years to performing electrophoresis separations in microfluidic format. Employing microfluidic technology for this purpose provides several benefits, such as improved transport control, reduced sample volumes, simplicity of operation, portability, greater accessibility, and reduced cost. The aim of this study is to develop microfluidic systems for high-performance separation of biochemical samples using electrophoresis methods. The first part of the thesis concerns the development of a fully integrated microfluidic chip for isoelectric focusing separation of proteins with whole-channel imaging detection. All the challenges posed in fabricating and integrating the chip were addressed. The chip was tested by performing protein and pI marker separations, and the separation results obtained from the chip were compared with those obtained from commercial cartridges. Side-by-side comparison of the results validated the developed chip and fabrication techniques. The research also focuses on improving the peak capacity and separation resolution of two counter-flow gradient electrofocusing methods: Temperature Gradient Focusing (TGF) and Micellar Affinity Gradient Focusing (MAGF). In these techniques, a temperature gradient across a microchannel or capillary is used to separate analytes. With an appropriate buffer, the temperature gradient creates a gradient in the electrophoretic velocity (TGF) or affinity (MAGF) of analytes and, if combined with a bulk counter-flow, ionic species concentrate at unique points where their total velocity is zero, and separate from each other. A bilinear temperature gradient is used along the separation channel to improve both peak capacity and separation resolution simultaneously. The temperature profile along the channel consists of a very sharp gradient used to pre-concentrate the sample, followed by a shallow gradient that increases separation resolution. A simple numerical model was applied to predict the improvement in resolution when using a bilinear gradient. A hybrid PDMS/glass chip integrated with planar micro-heaters for generating bilinear temperature gradients was fabricated using conventional sputtering and soft lithography techniques. A specialized design was developed for the heaters to achieve the desired bilinear profiles using both analytical and numerical modeling. To confirm the temperature profile along the channel, a two-color thermometry technique was also developed for measuring the temperature inside the chip. Separation performance was characterized by separating several different dyes, amino acids and peptides. Experiments showed a dramatic improvement in peak capacity and resolution of both techniques over the standard linear temperature gradients. Next, an analytical model was developed to investigate the effect of bilinear gradients in counter-flow gradient electrofocusing methods. The model provides a general equation for calculating the resolution for different gradients, diffusion coefficients and bulk flow scan rates. The results indicate that a bilinear gradient provides up to 100% improvement in separation resolution over the linear case. Additionally, for some scanning rates, an optimum bilinear gradient can be found that maximizes separation resolution. Numerical modeling was also developed to validate some of the results. The final part of the thesis describes the development of a two-dimensional separation system for protein separation, combining temperature gradient focusing (TGF) and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) in a PDMS/glass microfluidic chip. An experimental study was performed on separating a mixture of proteins using two characteristics: charge to mass ratio, and size. Experimental results showed a dramatic improvement in peak capacity over each of the one-dimensional separation techniques.

Humic-based organic compounds containing phenol or benzoic acid groups strongly compete with phosphates for specific binding sites on the surface of these colloidal particles. To study the interactions between phenol groups and the surface binding sites of unmodified or modified colloidal particles, chemical force spectrometry (CFS) was used as a tool to measure the adhesion force between an atomic force microscopy (AFM) tip terminated with a phenol self-assembled monolayer and colloidal particles under varying pH conditions. Two modification methods, co-precipitation and post-precipitation, were used to simulate the naturally-occurring phosphate and humic-acid adsorption process. The pH dependence of adhesion forces between phenol-terminated tip and colloidal particles could be explained by an interplay of electrostatic forces, the surface loading of the modifying phosphate or humic acid species and ionic hydrogen bonding. Polydimethylsiloxane (PDMS) is a widely-used polymer in microfluidic devices. PDMS surfaces are commonly modified to make it suitable for specific microfluidic devices. We studied the surface modification of PDMS using four perfluoroalkyl-triethoxysilane molecules of differing length of perfluorinated alkyl chain. The results show that the length of fluorinated alkyl chain has important effects on the density of surface modifying molecules, surface topography and surface zeta potential. The perfluorinated overlayer makes PDMS more efficient at supporting electroosmotic flow, which has potential applications in microfluidic devices. The kinetic study of RNase A, lysozyme C, α-lactalbumin and myoglobin at different concentrations adsorbed on the self-assembled monolayers of 1-octanethiol (OT-Au) and 1H, 1H, 2H, 2H-perfluorooctyl-1-thiol (FOT-Au) has been carried out. The results show a positive relationship between the lower protein concentration and the increased adsorption rate constant (ka) on both surfaces. At low concentrations, the protein adsorption on an OT-Au surface has greater ka than it on a FOT-Au surface. Comparing ka values for four proteins on OT-Au and FOT-Au surface demonstrates that hard proteins (lysozyme and RNase A) have larger ka than soft proteins (α-lactalbumin and myoglobin) on both surfaces. The discussion is based on the hydrophobicity of OT-Au and FOT-Au surfaces, as well as average superficial hydrophobicity, flexibility, size, stability, and surface induced conformation change of proteins. / Thesis (Ph.D, Chemistry) -- Queen's University, 2011-10-14 21:08:31.617