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Description
This work demonstrated a novel microfluidic device based on direct current (DC) insulator based dielectrophoresis (iDEP) for trapping individual mammalian cells in a microfluidic device. The novel device is also applicable for selective trapping of weakly metastatic mammalian breast cancer cells (MCF-7) from mixtures with mammalian Peripheral Blood Mononuclear Cells

This work demonstrated a novel microfluidic device based on direct current (DC) insulator based dielectrophoresis (iDEP) for trapping individual mammalian cells in a microfluidic device. The novel device is also applicable for selective trapping of weakly metastatic mammalian breast cancer cells (MCF-7) from mixtures with mammalian Peripheral Blood Mononuclear Cells (PBMC) and highly metastatic mammalian breast cancer cells, MDA-MB-231. The advantage of this approach is the ease of integration of iDEP structures in microfliudic channels using soft lithography, the use of DC electric fields, the addressability of the single cell traps for downstream analysis and the straightforward multiplexing for single cell trapping. These microfluidic devices are targeted for capturing of single cells based on their DEP behavior. The numerical simulations point out the trapping regions in which single cell DEP trapping occurs. This work also demonstrates the cell conductivity values of different cell types, calculated using the single-shell model. Low conductivity buffers are used for trapping experiments. These low conductivity buffers help reduce the Joule heating. Viability of the cells in the buffer system was studied in detail with a population size of approximately 100 cells for each study. The work also demonstrates the development of the parallelized single cell trap device with optimized traps. This device is also capable of being coupled detection of target protein using MALDI-MS.
ContributorsBhattacharya, Sanchari (Author) / Ros, Alexandra (Committee member) / Ros, Robert (Committee member) / Buttry, Daniel (Committee member) / Arizona State University (Publisher)
Created2013
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Description
The flow around a golf ball is studied using direct numerical simulation (DNS). An immersed boundary approach is adopted in which the incompressible Navier-Stokes equations are solved using a fractional step method on a structured, staggered grid in cylindrical coordinates. The boundary conditions on the surface are imposed using momentum

The flow around a golf ball is studied using direct numerical simulation (DNS). An immersed boundary approach is adopted in which the incompressible Navier-Stokes equations are solved using a fractional step method on a structured, staggered grid in cylindrical coordinates. The boundary conditions on the surface are imposed using momentum forcing in the vicinity of the boundary. The flow solver is parallelized using a domain decomposition strategy and message passing interface (MPI), and exhibits linear scaling on as many as 500 processors. A laminar flow case is presented to verify the formal accuracy of the method. The immersed boundary approach is validated by comparison with computations of the flow over a smooth sphere. Simulations are performed at Reynolds numbers of 2.5 × 104 and 1.1 × 105 based on the diameter of the ball and the freestream speed and using grids comprised of more than 1.14 × 109 points. Flow visualizations reveal the location of separation, as well as the delay of complete detachment. Predictions of the aerodynamic forces at both Reynolds numbers are in reasonable agreement with measurements. Energy spectra of the velocity quantify the dominant frequencies of the flow near separation and in the wake. Time-averaged statistics reveal characteristic physical patterns in the flow as well as local trends within dimples. A mechanism of drag reduction due to the dimples is confirmed, and metrics for dimple optimization are proposed.
ContributorsSmith, Clinton E (Author) / Squires, Kyle D (Thesis advisor) / Balaras, Elias (Committee member) / Herrmann, Marcus (Committee member) / Adrian, Ronald (Committee member) / Stanzione, Daniel C (Committee member) / Calhoun, Ronald (Committee member) / Arizona State University (Publisher)
Created2011
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Description
The evolution of single hairpin vortices and multiple interacting hairpin vortices are studied in direct numerical simulations of channel flow at Re-tau=395. The purpose of this study is to observe the effects of increased Reynolds number and varying initial conditions on the growth of hairpins and the conditions under which

The evolution of single hairpin vortices and multiple interacting hairpin vortices are studied in direct numerical simulations of channel flow at Re-tau=395. The purpose of this study is to observe the effects of increased Reynolds number and varying initial conditions on the growth of hairpins and the conditions under which single hairpins autogenerate hairpin packets. The hairpin vortices are believed to provide a unified picture of wall turbulence and play an important role in the production of Reynolds shear stress which is directly related to turbulent drag. The structures of the initial three-dimensional vortices are extracted from the two-point spatial correlation of the fully turbulent direct numerical simulation of the velocity field by linear stochastic estimation and embedded in a mean flow having the profile of the fully turbulent flow. The Reynolds number of the present simulation is more than twice that of the Re-tau=180 flow from earlier literature and the conditional events used to define the stochastically estimated single vortex initial conditions include a number of new types of events such as quasi-streamwise vorticity and Q4 events. The effects of parameters like strength, asymmetry and position are evaluated and compared with existing results in the literature. This study then attempts to answer questions concerning how vortex mergers produce larger scale structures, a process that may contribute to the growth of length scale with increasing distance from the wall in turbulent wall flows. Multiple vortex interactions are studied in detail.
ContributorsParthasarathy, Praveen Kumar (Author) / Adrian, Ronald (Thesis advisor) / Huang, Huei-Ping (Committee member) / Herrmann, Marcus (Committee member) / Arizona State University (Publisher)
Created2011
Description
Microfluidics is the study of fluid flow at very small scales (micro -- one millionth of a meter) and is prevalent in many areas of science and engineering. Typical applications include lab-on-a-chip devices, microfluidic fuel cells, and DNA separation technologies. Many of these microfluidic devices rely on micron-resolution velocimetry measurements

Microfluidics is the study of fluid flow at very small scales (micro -- one millionth of a meter) and is prevalent in many areas of science and engineering. Typical applications include lab-on-a-chip devices, microfluidic fuel cells, and DNA separation technologies. Many of these microfluidic devices rely on micron-resolution velocimetry measurements to improve microchannel design and characterize existing devices. Methods such as micro particle imaging velocimetry (microPIV) and micro particle tracking velocimetry (microPTV) are mature and established methods for characterization of steady 2D flow fields. Increasingly complex microdevices require techniques that measure unsteady and/or three dimensional velocity fields. This dissertation presents a method for three-dimensional velocimetry of unsteady microflows based on spinning disk confocal microscopy and depth scanning of a microvolume. High-speed 2D unsteady velocity fields are resolved by acquiring images of particle motion using a high-speed CMOS camera and confocal microscope. The confocal microscope spatially filters out of focus light using a rotating disk of pinholes placed in the imaging path, improving the ability of the system to resolve unsteady microPIV measurements by improving the image and correlation signal to noise ratio. For 3D3C measurements, a piezo-actuated objective positioner quickly scans the depth of the microvolume and collects 2D image slices, which are stacked into 3D images. Super resolution microPIV interrogates these 3D images using microPIV as a predictor field for tracking individual particles with microPTV. The 3D3C diagnostic is demonstrated by measuring a pressure driven flow in a three-dimensional expanding microchannel. The experimental velocimetry data acquired at 30 Hz with instantaneous spatial resolution of 4.5 by 4.5 by 4.5 microns agrees well with a computational model of the flow field. The technique allows for isosurface visualization of time resolved 3D3C particle motion and high spatial resolution velocity measurements without requiring a calibration step or reconstruction algorithms. Several applications are investigated, including 3D quantitative fluorescence imaging of isotachophoresis plugs advecting through a microchannel and the dynamics of reaction induced colloidal crystal deposition.
ContributorsKlein, Steven Adam (Author) / Posner, Jonathan D (Thesis advisor) / Adrian, Ronald (Committee member) / Chen, Kangping (Committee member) / Devasenathipathy, Shankar (Committee member) / Frakes, David (Committee member) / Arizona State University (Publisher)
Created2011
Description
DNA and DNA nanoassemblies such as DNA origamis have large potential in biosensing, drug delivery, nanoelectronic circuits, and biological computing requiring suitable methods for migration and precise positioning. Insulator-based dielectrophoresis (iDEP) provides an efficient and matrix-free approach for manipulation of micro-and nanometer-sized objects. In order to exploit iDEP for naturally

DNA and DNA nanoassemblies such as DNA origamis have large potential in biosensing, drug delivery, nanoelectronic circuits, and biological computing requiring suitable methods for migration and precise positioning. Insulator-based dielectrophoresis (iDEP) provides an efficient and matrix-free approach for manipulation of micro-and nanometer-sized objects. In order to exploit iDEP for naturally formed DNA and DNA nanoassemblies, a detailed understanding of the underlying polarization and dielectrophoretic migration is essential. The shape and the counterion distribution are considered two essential factors in the polarization mechanism. Here, the dielectrophoretic behavior of 6-helix bundle (6HxB) and triangle DNA origamis with identical sequences but substantial topological differences was explored. The polarizability models were discussed for the two species according to their structural difference. The experimental observations reveal distinct iDEP trapping behavior in low frequency AC electric fields in addition to numerical simulations showing a considerable contribution of the electrophoretic transport of the DNA origami species in the DEP trapping regions. Furthermore, the polarizabilities of the two species were determined by measuring the migration times through a potential landscape exhibiting dielectrophoretic barriers. The resulting migration times correlate to the depth of the dielectrophoretic potential barrier and the escape characteristics of the DNA origamis according to an adapted Kramer’s rate model. The orientations of both species in the escape process were studied. Finally, to study the counterion distribution around the DNA molecules, both λ-DNA and 6HxB DNA were used in a phosphate buffer containing magnesium, revealing distinctive negative dielectrophoretic trapping behavior as opposed to positive trapping in a sodium/potassium phosphate buffer system.
ContributorsGan, Lin (Author) / Ros, Alexandra (Thesis advisor) / Buttry, Daniel (Committee member) / Liu, Yan (Committee member) / Arizona State University (Publisher)
Created2015
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Description
This work helps to explain the drag reduction mechanisms at low and moderate turbulent Reynolds numbers in pipe flows. Through direct numerical simulation, the effects of wall oscillations are observed on the turbulence in both the near wall and the bulk region. Analysis of the average Reynolds

This work helps to explain the drag reduction mechanisms at low and moderate turbulent Reynolds numbers in pipe flows. Through direct numerical simulation, the effects of wall oscillations are observed on the turbulence in both the near wall and the bulk region. Analysis of the average Reynolds Stresses at various phases of the flow is provided along with probability density functions of the fluctuating components of velocity and vorticity. The flow is also visualized to observe, qualitatively, changes in the total and fluctuating field of velocity and vorticity. Linear Stochastic Estimation is used to create a conditional eddy (associated with stress production) in the flow and visualize the effects of transverse wall oscillations on hairpin growth, auto-generation and structure.
ContributorsCoxe, Daniel (Author) / Peet, Yulia (Thesis advisor) / Adrian, Ronald (Thesis advisor) / Herrmann, Marcus (Committee member) / Arizona State University (Publisher)
Created2019
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Description
Disease prevention and personalized treatment will be impacted by the continued integration of protein biomarkers into medical practice. While there are already numerous biomarkers used clinically, the detection of protein biomarkers among complex matrices remains a challenging problem. One very important strategy for improvements in clinical application of biomarkers is

Disease prevention and personalized treatment will be impacted by the continued integration of protein biomarkers into medical practice. While there are already numerous biomarkers used clinically, the detection of protein biomarkers among complex matrices remains a challenging problem. One very important strategy for improvements in clinical application of biomarkers is separation/preconcentration, impacting the reliability, efficiency and early detection. Electrophoretic exclusion can be used to separate, purify, and concentrate biomarkers. This counterflow gradient technique exploits hydrodynamic flow and electrophoretic forces to exclude, enrich, and separate analytes. The development of this technique has evolved onto an array-based microfluidic platform which offers a greater range of geometries/configurations for optimization and expanded capabilities and applications. Toward this end of expanded capabilities, fundamental studies of subtle changes to the entrance flow and electric field configurations are investigated. Three closely related microfluidic interfaces are modeled, fabricated and tested. A charged fluorescent dye is used as a sensitive and accurate probe to test the concentration variation at these interfaces. Models and experiments focus on visualizing the concentration profile in areas of high temporal dynamics, and show strong qualitative agreement, which suggests the theoretical assessment capabilities can be used to faithfully design novel and more efficient interfaces. Microfluidic electrophoretic separation technique can be combined with electron microscopy as a protein concentration/purification step aiding in sample preparation. The integrated system with grids embedded into the microdevice reduces the amount of time required for sample preparation to less than five minutes. Spatially separated and preconcentrated proteins are transferred directly from an upstream reservoir onto grids. Dilute concentration as low as 0.005 mg/mL can be manipulated to achieve meaningful results. Selective concentration of one protein from a mixture of two proteins is also demonstrated. Electrophoretic exclusion is also used for biomarker applications. Experiments using a single biomarker are conducted to assess the ability of the microdevice for enrichment in central reservoirs. A mixture of two protein biomarkers are performed to evaluate the proficiency of the device for separations capability. Moreover, a battery is able to power the microdevice, which facilitates the future application as a portable device.
ContributorsZhu, Fanyi (Author) / Hayes, Mark (Thesis advisor) / Ros, Alexandra (Committee member) / Buttry, Daniel (Committee member) / Arizona State University (Publisher)
Created2019
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Description
X-ray crystallography is the most widely used method to determine the structure of proteins, providing an understanding of their functions in all aspects of life to advance applications in fields such as drug development and renewable energy. New techniques, namely serial femtosecond crystallography (SFX), have unlocked the ability to unravel

X-ray crystallography is the most widely used method to determine the structure of proteins, providing an understanding of their functions in all aspects of life to advance applications in fields such as drug development and renewable energy. New techniques, namely serial femtosecond crystallography (SFX), have unlocked the ability to unravel the structures of complex proteins with vital biological functions. A key step and major bottleneck of structure determination is protein crystallization, which is very arduous due to the complexity of proteins and their natural environments. Furthermore, crystal characteristics govern data quality, thus need to be optimized to attain the most accurate reconstruction of the protein structure. Crystal size is one such characteristic in which narrowed distributions with a small modal size can significantly reduce the amount of protein needed for SFX. A novel microfluidic sorting platform was developed to isolate viable ~200 nm – ~600 nm photosystem I (PSI) membrane protein crystals from ~200 nm – ~20 μm crystal samples using dielectrophoresis, as confirmed by fluorescence microscopy, second-order nonlinear imaging of chiral crystals (SONICC), and dynamic light scattering. The platform was scaled-up to rapidly provide 100s of microliters of sorted crystals necessary for SFX, in which similar crystal size distributions were attained. Transmission electron microscopy was used to view the PSI crystal lattice, which remained well-ordered postsorting, and SFX diffraction data was obtained, confirming a high-quality, viable crystal sample. Simulations indicated sorted samples provided accurate, complete SFX datasets with 3500-fold less protein than unsorted samples. Microfluidic devices were also developed for versatile, rapid protein crystallization screening using nanovolumes of sample. Concentration gradients of protein and precipitant were generated to crystallize PSI, phycocyanin, and lysozyme using modified counterdiffusion. Additionally, a passive mixer was created to generate unique solution concentrations within isolated nanowells to crystallize phycocyanin and lysozyme. Crystal imaging with brightfield microscopy, UV fluorescence, and SONICC coupled with numerical modeling allowed quantification of crystal growth conditions for efficient phase diagram development. The developed microfluidic tools demonstrated the capability of improving samples for protein crystallography, offering a foundation for continued development of platforms to aid protein structure determination.
ContributorsAbdallah, Bahige G (Author) / Ros, Alexandra (Thesis advisor) / Buttry, Daniel (Committee member) / Hayes, Mark (Committee member) / Arizona State University (Publisher)
Created2016
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Description
Portable devices rely on battery systems that contribute largely to the overall device form factor and delay portability due to recharging. Membraneless microfluidic fuel cells are considered as the next generation of portable power sources for their compatibility with higher energy density reactants. Microfluidic fuel cells are potentially cost effective

Portable devices rely on battery systems that contribute largely to the overall device form factor and delay portability due to recharging. Membraneless microfluidic fuel cells are considered as the next generation of portable power sources for their compatibility with higher energy density reactants. Microfluidic fuel cells are potentially cost effective and robust because they use low Reynolds number flow to maintain fuel and oxidant separation instead of ion exchange membranes. However, membraneless fuel cells suffer from poor efficiency due to poor mass transport and Ohmic losses. Current microfluidic fuel cell designs suffer from reactant cross-diffusion and thick boundary layers at the electrode surfaces, which result in a compromise between the cell's power output and fuel utilization. This dissertation presents novel flow field architectures aimed at alleviating the mass transport limitations. The first architecture provides a reactant interface where the reactant diffusive concentration gradients are aligned with the bulk flow, mitigating reactant mixing through diffusion and thus crossover. This cell also uses porous electro-catalysts to improve electrode mass transport which results in higher extraction of reactant energy. The second architecture uses porous electrodes and an inert conductive electrolyte stream between the reactants to enhance the interfacial electrical conductivity and maintain complete reactant separation. This design is stacked hydrodynamically and electrically, analogous to membrane based systems, providing increased reactant utilization and power. These fuel cell architectures decouple the fuel cell's power output from its fuel utilization. The fuel cells are tested over a wide range of conditions including variation of the loads, reactant concentrations, background electrolytes, flow rates, and fuel cell geometries. These experiments show that increasing the fuel cell power output is accomplished by increasing reactant flow rates, electrolyte conductivity, and ionic exchange areas, and by decreasing the spacing between the electrodes. The experimental and theoretical observations presented in this dissertation will aid in the future design and commercialization of a new portable power source, which has the desired attributes of high power output per weight and volume and instant rechargeability.
ContributorsSalloum, Kamil S (Author) / Posner, Jonathan D (Thesis advisor) / Adrian, Ronald (Committee member) / Christen, Jennifer (Committee member) / Phelan, Patrick (Committee member) / Chen, Kangping (Committee member) / Arizona State University (Publisher)
Created2010
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Description
Locomotion of microorganisms is commonly observed in nature. Although microorganism locomotion is commonly attributed to mechanical deformation of solid appendages, in 1956 Nobel Laureate Peter Mitchell proposed that an asymmetric ion flux on a bacterium's surface could generate electric fields that drive locomotion via self-electrophoresis. Recent advances in nanofabrication have

Locomotion of microorganisms is commonly observed in nature. Although microorganism locomotion is commonly attributed to mechanical deformation of solid appendages, in 1956 Nobel Laureate Peter Mitchell proposed that an asymmetric ion flux on a bacterium's surface could generate electric fields that drive locomotion via self-electrophoresis. Recent advances in nanofabrication have enabled the engineering of synthetic analogues, bimetallic colloidal particles, that swim due to asymmetric ion flux originally proposed by Mitchell. Bimetallic colloidal particles swim through aqueous solutions by converting chemical fuel to fluid motion through asymmetric electrochemical reactions. This dissertation presents novel bimetallic motor fabrication strategies, motor functionality, and a study of the motor collective behavior in chemical concentration gradients. Brownian dynamics simulations and experiments show that the motors exhibit chemokinesis, a motile response to chemical gradients that results in net migration and concentration of particles. Chemokinesis is typically observed in living organisms and distinct from chemotaxis in that there is no particle directional sensing. The synthetic motor chemokinesis observed in this work is due to variation in the motor's velocity and effective diffusivity as a function of the fuel and salt concentration. Static concentration fields are generated in microfluidic devices fabricated with porous walls. The development of nanoscale particles that swim autonomously and collectively in chemical concentration gradients can be leveraged for a wide range of applications such as directed drug delivery, self-healing materials, and environmental remediation.
ContributorsWheat, Philip Matthew (Author) / Posner, Jonathan D (Thesis advisor) / Phelan, Patrick (Committee member) / Chen, Kangping (Committee member) / Buttry, Daniel (Committee member) / Calhoun, Ronald (Committee member) / Arizona State University (Publisher)
Created2011