Biomedical Engineering and Bioengineering Commons^™

Microfluidic Electrical Impedance Spectroscopy System Automation And Characterization, Keaton Frahmann

Master's Theses

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 …

A Protocol For Achieving Adherent Cell Culture Within A Microfluidic Device, Tarra Danielle Sanders

Master's Theses

The goal of this study is to design a protocol for the adherent cell culture within a novel microfluidic device. Microscale cell culture protocols were developed for loading cells using poly-L-lysine to enhance adherent cell culture of murine derived NIH 3T3 fibroblasts. This work sought to develop a method for adherent microculture by examining various sterilization, surface treatment, and seeding techniques. Using a vacuum suction loading technique, air plasma treatment and a poly-L-lysine surface treatment adherent cell culture was observed within the device. The work presented here is part of a collaborative effort that aims to develop protocols for the …

Opendrop Elisa: Surface Development For Digital Microfluidic Elisa Testing, Michaela Q. Mead, Madeline G. Jackson, Jacob G. Taylor

Biomedical Engineering

This project aimed to develop a process to create a hydrophobic coating with capture antibodies attached. This process was developed in an attempt to create a digital microfluidic platform upon which an enzyme-linked immunosorbent assay (ELISA) could be run, indicated for clinicians and educators in low-resource settings. Four requirements dictated the course of the design process: 1) the contact angle with water of the hydrophobic layer must be greater than 90º, 2) the colorimetric response of a positive ELISA result on our platform must be at least 85% of that run on polystyrene with the same result, 3) the platform …

Insulative (Direct Current) Dielectrophoretic Foul-Less Filtration In Microfuidic Systems, Matthew A A Whitman

Master's Theses

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 …

Fluid Delivery System For A Cell Culture On A Microfluidic Chip, Austin J. Roeder, Colleen A. Richards, Emily A. Matteson

Biomedical Engineering

This project report provides a description of the progress made in the development of a fluid delivery system for a microfluidic cell culture on a chip. The system is intended to be used in a humidified incubator in a university laboratory and the fluid delivery system is required to exist and operate within that incubator for extended periods of time. Therefore, the system will be gravity-driven and contain no electronic components. The key specification of the system is to provide fluid flow at a constant velocity.

After manufacturing and testing the device, all specifications were met except for the fluid …

Microfluidic Electrical Impedance Spectroscopy, John J. Foley

Master's Theses

The goal of this study is to design and manufacture a microfluidic device capable of measuring changes in impedance valuesof microfluidic cell cultures. Tocharacterize this, an interdigitated array of electrodes was patterned over glass, where it was then bonded to a series of fluidic networks created in PDMS via soft lithography. The device measured ethanol impedance initially to show that values remain consistent over time. Impedance values of water and 1% wt. saltwater were compared to show that the device is able to detect changes in impedance, with up to a 60% reduction in electrical impedance in saltwater. Cells were …

Poly Drop, Zachary T. Scott, Lilly J. Paul

Computer Engineering

Poly Drop is a software interface to control an Open Drop digital micro-fluidics system. We obtained a hardware system from Gaudi labs. Our task was to create a Graphical User Interface that made the control of the device easier and more automated for better testing. We created software that had 3 parts: a control GUI, arduino code to control the hardware, and Image Analysis that gives the user information such as location and color of liquid drops as they move across the electrode grid of the Open Drop system. The GUI was developed using Java Swing. The communication between the …

Functional 3-D Cellulose And Nitrocellulose Paper-Based, Microfluidic Device Utilizing Elisa Technology For The Detection/Distinction Between Hemorrhagic And Ischemic Strokes, Alicia Leanne Holler

Master's Theses

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 …

Development, Characterization, And Optimization Of Process For Minimal Functional Barrier And Channel Width In Nitrocellulose Paper Microfluidic Diagnostic Platforms, Ryan Daniel Silva

Biomedical Engineering

A central goal of diagnostic microfluidics is to reduce the cost of diagnostic medicine by reducing the equipment and reagents needed to perform diagnostic tests. The literature has demonstrated that a wax printer can be used to pattern nitrocellulose paper with hydrophobic barriers to direct a sample in a defined reaction path, eliminating the need for external pumps and controllers. However, manufacturing methods for minimizing sample volume (and thus reagent volume) in a paper-diagnostic chip have not been well defined. In this work, we experimentally determine manufacturing processes for creating functional features of minimal size—effectively reducing the sample size and …

An Analysis Of Eliminating Electroosmotic Flow In A Microfluidic Pdms Chip, Cecile D. Redington

Master's Theses

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 …

The Fabrication & Characterization Of An Electrokinetic Microfluidic Pump From Su-8, A Negative Epoxy-Based Photoresist, Nash Anderson

Master's Theses

Microfluidics refers to manipulation, precise control, and behavior of fluids at the micro and nanoliter scales. It has entered the realm of science as a way to precisely measure or mix small amounts of fluid to perform highly controlled reactions. Glass and polydimethylsiloxane (PDMS) are common materials used to create microfluidic devices; however, glass is difficult to process and PDMS is relatively hydrophobic. In this study, SU-8, an epoxy based (negative) photoresist was used to create various electrokinetic microfluidic chips. SU-8 is commonly used in microelectromechanical design. Spin coating of various SU-8 formulations allows for 1 μm to 100 μm …

A Microfluidic Pretreatment Isoelectric Focusing Dc-Field Assisted H-Filter For The Separation Of Charged Particles, Cameron Sean Carroll

Biomedical Engineering

Microfluidic devices are small portable diagnostic chips that allow for analysis of a biologic sample at relatively low cost. This makes them ideal for settings where a hospital is unavailable. The microfluidic pretreatment isoelectric focusing DC-field assisted H-filter is a device that allows for the separation of differently charged particles in a biologic sample. It does this by employing the concepts of diffusion, electrophoresis, and isoelectric focusing. This is perfect for isolation of a desired analyte or separation of waste from a sample in order to achieve better diagnostic results. The device was first modeled mathematically and visually using Comsol. …

Hydrodynamic Focused Passive Separation Under Continuous Flow In A Microfluidic Chip, Jad Kanbar

Master's Theses

A continuous flow, passive separation device was designed using an equivalent circuit to create variable flow rates for hydrodynamic focusing to drain channels and collection outlets. By varying the diameter of the sample inlet connection into the reservoir, the particle position was influenced significantly, which enabled desired separations. Additionally it was noted that the relative, horizontal position of the inlet also had a significant influence on particle position within the device. A dimensionless number, the Characteristic Sample Inlet, was developed to relate geometric properties of the inlet reservoir to downstream particle distribution. It was found that a 2:1 ratio between …

Impedance Sensing Of N2a And Astrocytes As Grounds For A Central Nervous System Cancer Diagnostic Device, Fraser Traves Smith Grove

Master's Theses

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 …

Integration Of Microfluidics With Surface Plasmon Resonance, Scott B. Fratzke

Master's Theses

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 …

Design, Fabrication, And Implementation Of A Single-Cell Capture Chamber For A Microfluidic Impedance Sensor, Joshua-Jed Doria Fadriquela

Master's Theses

A microfluidic device was created for single-cell capture and analysis using polydimethylsiloxane (PDMS) channels and a glass substrate to develop a microfluidic single-cell impedance sensor for cell diagnostics. The device was fabricated using photolithography to create a master mold which in turn will use soft lithography to create the PDMS components for constant device production. The commercial software, COMSOLTM Multiphysics, was used to quantify the fluid dynamics in shallow micro-channels.

The device will be able to capture a cell and sequester it long enough to enable measurement of the impedance spectra that can characterize cell. The proposed device will be …