This thesis successfully integrates laminate microfluidic devices with an analytic Surface Plasmon Resonance (SPR) instrument. Integration was accomplished at low-cost using materials such as polydimethylsiloxane (PDMS), Poly(methyl methacrylate) (PMMA), Tygon tubing, and a 3-way stopcock. The main components of this thesis are the design and fabrication of the low-cost, in-house fluidics that can integrate with upstream microfluidics and the validation of the in-house fluidics using the Biosensing Instruments BI-2000 SPR instrument. The low-cost fluidics was designed and fabricated “in-house” using a novel investment casting technique that required the use of laser cutting technology to make a master cast, and candle wax to make the fluidic flow gasket. Integration of upstream microfluidic devices is the next step towards fully integrated point-of-care (POC) diagnostics. Development of low-cost POC diagnostics will enable physicians to diagnosis patients outside of clinical settings, granting treatment access to a much wider population. Surface Plasmon Resonance is used for its detection abilities combined with its ability to perform real-time sample analysis. Validation of the in-house fluidics was accomplished by conducting (2) experiments: (1) to compare the angular shift elicited by ethanol solutions between in-house fluidics, factory fluidics, and the literature, and (2) to compare the angular shift between in-house fluidics and factory fluidics caused by the cleaving of fibroblasts from the SPR sensor chip. Successful comparisons made in both experiments proved successful development of low-cost fluidics that could integrate upstream microfluidic devices.

Filtration is a technology that is used almost ubiquitously in society from uses raging from filtration of macroparticles from water to pharmaceutical grade filtration products to remove anything larger than a protein. However, with such a wide range of uses, most filtration products have the same issue; membrane clogging (fouling) that prevents continuous use and requires frequent maintenance. This thesis hypothesizes that by applying a direct current (DC) to an insulating array of posts, they will create a foul-less insulative dielectrophoretic filter (iDEP) that does not clog since particles will levitate above the insulating array. This thesis tested an inherited device (legacy device) and found that its design did not perform the desired foul-less filtration operation under the tested conditions. Therefore, using COMSOL simulations, the conditions of testing and improved deign were developed to fruition. These devices were fabricated and tested and found to successfully levitate yeast particles above the foul-less filtration array using a direct current insulative dielectrophoretic (iDEP) filter. Additionally, different post geometries were observed and how they affect the dielectric force on particles. Although a foul-less filter was not successfully developed over the course of this thesis, the groundwork for development of DC iDEP has been set.

3-D paper-based microfluidic devices (micoPADs) are small and portable devices made out of paper that offer a promising platform for diagnostic applications outside of a laboratory. These devices are easy to use, low cost, require no power source, and capable of detecting multiple targets simultaneously. The work in this thesis demonstrated the ability of a 3-D paper-microfluidic platform to simultaneously detect 5 targets. Rubber cord stock was used in conjunction with an acrylic housing unit to apply pressure along the edge of the channel. The indirect pressure application was successful in promoting vertical fluid flow between layers. Average channel development times were recorded between 110 seconds and 150 seconds. The implementation of the 3-D paper-microfluidic platform as a diagnostic device was validated with a colorimetric glucose assay. In a novel application, reagents were deposited onto the 3-D platform via a glucose reagent pencil created by Martinez et al. A visual signal was observed for the successful detection of glucose at a concentration of 1.2 mM. These results offer promise for future work in combing new reagent deposition techniques with a multi-layer paper-microfluidic platform. Overall, this research made advancements in the design of a paper-microfluidic platform capable of the simultaneous detection of 5 targets.

In this work, a novel microfluidic cell culture platform capable of automated electrical impedance measurements and immunofluorescence and brightfield microscopy was developed for further in-vitro cellular research intended to optimize cell culture conditions. The microfluidic system design, fabrication, automation, and design verification testing are described. Electrical and optical measurements of the 16 parallel cell culture chambers were automated via a custom LabView interface. A proposed design change will enable gas diffusion, removing the need for an environmental enclosure and allow long-term cell culture experiments. This "lab on a chip" system miniaturizes and automates experiments improving testing throughput and accuracy while creating a highly controllable microenvironment for cell culture. Such a system can be applied to drug development, bioassays, diagnostics, and animal testing alternatives. This work is part of a collaborative effort to define protocols for the electrical and optical characterization of cell culture within a novel microfluidic device with the intent of optimizing microenvironment conditions.

This thesis encompasses a feasibility study of using low-cost materials to manufacture microfluidic chips that can perform the same functions as chips manufactured using traditional methods within an acceptable range of efficiency of chips created with more exotic methods and materials. The major parts of the project are the selection and characterization of the fabrication methods for creating the channels for fluid flow, the methods for sealing the channels to create a usable chip and the electrophoretic separations of carboxylated microspheres of different potentials. In this work we seek to answer the question if laser-etched PMMA microfluidic chips are comparable in functionality to microfluidic chips created with PDMS or glass. In the process of answering this question we will touch on FEA modeling, characterization of the manufacturing process and multiple prototype designs while keeping within the low-cost theme. The purpose of capillary electrophoresis is to separate proteins based on their inherent electric charge. Capillary electrophoresis is a standard chip design used in the microfluidics world to prove a new fabrication method or chip material before branching out to other experiments because it is a fairly simple and robust design. Common problems associated with the manufacturing methods and materials were taken into account such as electroosmotic flow and chip sealing. CZE designs from literature were referenced to create a chip that would separate carboxylated microbeads with reasonable resolution. Wire electrodes were affixed to the chip to induce electric fields for the electrophoresis experiments. The goal of this thesis is to prove the manufacturing methods and attain results within 70% of literature standards.

The purpose of this thesis project is to demonstrate and evaluate an enzyme-linked immunosorbent assay (ELISA) on a paper microfluidic device platform. The integration of ELISA technology onto paper microfluidic chips allows for a quantitative detection of stroke biomarkers, such as glial fibrillary acidic protein (GFAP). Dye experiments were performed to confirm fluid connectivity throughout the 3D chips. Several chip and housing designs were fabricated to determine an optimal design for the microfluidic device. Once this design was finalized, development time testing was performed. The results confirmed that the paper microfluidic device could successfully route fluid throughout its channels at a reasonable rate. For the biochemistry portion of this thesis project, antibodies were selected to target the intended stroke biomarker: GFAP. However, due to antibody pairing complications, the protein chosen for this project was natural human cardiac troponin T, which is elevated in the bloodstream of patients who have suffered a stroke. Several antibody experiments were performed to help finalize the procedure for performing an ELISA on the paper chip. The final antibody experiment was able to demonstrate that a paper microfluidic device utilizing ELISA techniques can successfully detect a stroke biomarker at physiologically relevant concentrations. Overall, this project supported the ability to accurately and effectively diagnose stroke in a timely manner through the use of a paper microfluidic device.

The goal of this project is to eliminate electroosmotic flow (EOF) in a microfluidic chip. EOF is a naturally occurring phenomenon at the fluid-surface interface in microfluidic chips when an electric field is applied across the fluid. When isoelectric focusing (IEF) is carried out to separate proteins based on their surface charge, the analytes must remain in the separation chamber, and not migrate to adjacent features in the microfluidic chip, which happens with EOF. For this project, a microfluidic chip was designed and commissioned to be photolithographically transferred onto a Si wafer. A PDMS component was then casted on the Si wafer and plasma bonded to a glass substrate. This chip was initially designed to carry out continuous IEF, and the focus of the project was shifted to the analysis of eliminating EOF in a microfluidic chamber. Per previous research test methods, methylcellulose will be used to analyze the phenomenon of electroosmotic flow in the chamber. A COMSOL model is used a theoretical basis of comparison when analyzing the flow velocities of the treated versus untreated microfluidic chips. The purpose of this project is to use the research performed in on this chip as a precursor to future analyses of continuous IEF on microfluidic chips in the Cal Poly Microfluidics group.

This thesis utilizes previously described manufacturing and design techniques for the creation of a PDMS-glass bonded microfluidic device, capable of electrochemical impedance spectroscopy (EIS). EIS has been used across various fields of research for different diagnostic needs. The major aim of this thesis was to capture cancerous and non-cancerous cells between micron sized electrodes within a microfluidic path, and to complete analysis on the measured impedances recorded from the two differing cell types. Two distinct ranges of impedance frequency were analyzed – the α dispersion range, which quantifies the impedance of the membranes of the cells of interest, and the β dispersion range, which quantifies the impedance of the cytosol of the cells of interest. This thesis is unique in the fact that it looks at the cellular impedances of two types of neural cells, which has not been documented previously in literature. The type of cancerous cells analyzed were Neuro-2-A cells, an immortalized line of murine glio/neuroblastoma. The type of non-cancerous cells analyzed were murine primary astrocytes, a mortal line of neurological support cells found throughout the nervous system, and with great abundance in the brain. By using a LabView program coded by a previous Cal Poly student, a sweep scan across a wide frequency range was completed on both cell types, and statistical analysis was completed on target frequencies of interest. A significant difference was found between the two cell lines’ membrane impedances, however no difference was found between the cytoplasm impedances. In total, this thesis aimed to fabricate a reusable microfluidic device capable of EIS for future Cal Poly students, create a protocol suitable for cell culturing and device operation, and to lay a foundation of knowledge for impedance comparisons regarding neural cancerous and non-cancerous cells.

Understanding cell migration is a key step in unraveling many physiological phenomena and predicting several pathologies, such as cancer metastasis. In particular, confinement has been proven to be a key factor in the cellular migration strategy choice. As our insight in the field improves, new tools are needed in order to empower biologists’ analysis capabilities. In this framework, microfluidic devices have been used to engineer the mechanical and spatial stimuli and to investigate cellular migration response in a more controlled way. In this work, we will review the existing technologies employed in the realization of microfluidic cellular migration assays, namely the soft lithography of PDMS and hydrogels and femtosecond laser micromachining. We will give an overview of the state of the art of these devices, focusing on the different geometrical configurations that have been exploited to study specific aspects of cellular migration. Our scope is to highlight the advantages and possibilities given by each approach and to envisage the future developments in in vitro migration studies under spatial confinement in microfluidic devices.

This work presents the development of 3D printed microfluidic devices and their application to microchip analysis. Initial work was focused on the development of the printer resin as well as the development of the general rules for resolution that can be achieved with stereolithographic 3D printing. The next stage of this work involved the characterization of the printer with a variety of interior and exterior resolution features. I found that the minimum positive and negative feature sizes were about 20 μm in either case. Additionally, micropillar arrays were printed with pillar diameters as small as 16 μm. To demonstrate one possible application of these small resolution features I created microfluidic bead traps capable of capturing 25 μm polystyrene particles as a step toward capturing cells. A second application which I pioneered was the creation of devices for microchip electrophoresis. I separated 3 preterm birth biomarkers with good resolution (2.1) and efficiency (3600 plates), comparable to what has been achieved in conventionally fabricated devices. Lastly, I have applied some of the unique capabilities of our 3D printer to a variety of other device applications through collaborative projects. I have created microchips with a natural masking monolith polymerization window, spiral electrodes for capacitively coupled contactless conductivity detection, and a removable electrode insert chip. This work demonstrates the ability to 3D print microfluidic structures and their application to a variety of analyses.