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Description
Cellular heterogeneity is a key factor in various cellular processes as well as in disease development, especially associated with immune response and cancer progression. Cell-to-cell variability is considered to be one of the major obstacles in early detection and successful treatment of cancer. Most present technologies are based on bulk cell analysis, which results in averaging out the results acquired from a group of cells and hence missing important information about individual cells and their behavior. Understanding the cellular behavior at the single-cell level can help in obtaining a complete profile of the cell and to get a more in-depth knowledge of cellular processes. For example, measuring transmembrane fluxes oxygen can provide a direct readout of the cell metabolism.
The goal of this thesis is to design, optimize and implement a device that can measure the oxygen consumption rate (OCR) of live single cells. A microfluidic device has been designed with the ability to rapidly seal and unseal microchambers containing individual cells and an extracellular optical oxygen sensor for measuring the OCR of live single cells. The device consists of two parts, one with the sensor in microwells (top half) and the other with channels and cells trapped in Pachinko-type micro-traps (bottom half). When the two parts of the device are placed together the wells enclose each cell. Oil is flown in through the channels of the device to produce isolated and sealed microchamber around each cell. Different fluids can be flowed in and out of the device, alternating with oil, to rapidly switch between sealed and unsealed microenvironment around each cell. A fluorescent ratiometric dual pH and oxygen sensor is placed in each well. The thesis focuses on measuring changes in the oxygen consumption rate of each cell within a well. Live and dead cells are identified using a fluorescent live/dead cell assay. Finally, the technology is designed to be scalable for high-throughput applications by controlling the flow rate of the system and increasing the cell array density.
The goal of this thesis is to design, optimize and implement a device that can measure the oxygen consumption rate (OCR) of live single cells. A microfluidic device has been designed with the ability to rapidly seal and unseal microchambers containing individual cells and an extracellular optical oxygen sensor for measuring the OCR of live single cells. The device consists of two parts, one with the sensor in microwells (top half) and the other with channels and cells trapped in Pachinko-type micro-traps (bottom half). When the two parts of the device are placed together the wells enclose each cell. Oil is flown in through the channels of the device to produce isolated and sealed microchamber around each cell. Different fluids can be flowed in and out of the device, alternating with oil, to rapidly switch between sealed and unsealed microenvironment around each cell. A fluorescent ratiometric dual pH and oxygen sensor is placed in each well. The thesis focuses on measuring changes in the oxygen consumption rate of each cell within a well. Live and dead cells are identified using a fluorescent live/dead cell assay. Finally, the technology is designed to be scalable for high-throughput applications by controlling the flow rate of the system and increasing the cell array density.
ContributorsRodrigues, Meryl (Author) / Meldrum, Deirdre (Thesis advisor) / Kelbauskas, Laimonas (Committee member) / LaBelle, Jeffrey (Committee member) / Arizona State University (Publisher)
Created2014
Description
In the search for chemical biosensors designed for patient-based physiological applications, non-invasive diagnostic approaches continue to have value. The work described in this thesis builds upon previous breath analysis studies. In particular, it seeks to assess the adsorptive mechanisms active in both acetone and ethanol biosensors designed for breath analysis. The thermoelectric biosensors under investigation were constructed using a thermopile for transduction and four different materials for biorecognition. The analytes, acetone and ethanol, were evaluated under dry-air and humidified-air conditions. The biosensor response to acetone concentration was found to be both repeatable and linear, while the sensor response to ethanol presence was also found to be repeatable. The different biorecognition materials produced discernible thermoelectric responses that were characteristic for each analyte. The sensor output data is presented in this report. Additionally, the results were evaluated against a mathematical model for further analysis. Ultimately, a thermoelectric biosensor based upon adsorption chemistry was developed and characterized. Additional work is needed to characterize the physicochemical action mechanism.
ContributorsWilson, Kimberly (Author) / Guilbeau, Eric (Thesis advisor) / Pizziconi, Vincent (Thesis advisor) / LaBelle, Jeffrey (Committee member) / Arizona State University (Publisher)
Created2011
Description
Volume depletion can lead to migraines, dizziness, and significant decreases in a subject's ability to physically perform. A major cause of volume depletion is dehydration, or loss in fluids due to an imbalance in fluid intake to fluid excretion. Because proper levels of hydration are necessary in order to maintain both short and long term health, the ability to monitor hydration levels is growing in clinical demand. Although devices capable of monitoring hydration level exist, these devices are expensive, invasive, or inaccurate and do not offer a continuous mode of measurement. The ideal hydration monitor for consumer use needs to be characterized by its portability, affordability, and accuracy. Also, this device would need to be noninvasive and offer continuous hydration monitoring in order to accurately assess fluctuations in hydration data throughout a specified time period. One particular method for hydration monitoring that fits the majority of these criteria is known as bioelectric impedance analysis (BIA). Although current devices using BIA do not provide acceptable levels of accuracy, portability, or continuity in data collection, BIA could potentially be modified to fit many, if not all, desired customer specifications. The analysis presented here assesses the viability of using BIA as a new standard in hydration level measurement. The analysis uses data collected from 22 subjects using an existing device that employs BIA. A regression derived for estimating TBW based on the parameters of age, weight, height, sex, and impedance is presented. Using impedance data collected for each subject, a regression was also derived for estimating impedance based on the factors of age, weight, height, and sex. The derived regression was then used to calculate a new impedance value for each subject, and these new impedance values were used to estimate TBW. Through a paired-t test between the TBW values derived by using the direct measurements versus the calculated measurements of impedance, the two samples were found to be comparable. Considerations for BIA as a noninvasive measurement of hydration are discussed.
ContributorsTenorio, Jorge Antonio (Author) / LaBelle, Jeffrey (Thesis director) / Pizziconi, Vincent (Committee member) / Spano, Mark (Committee member) / Barrett, The Honors College (Contributor) / W. P. Carey School of Business (Contributor) / Harrington Bioengineering Program (Contributor)
Created2013-05
Description
The action/adventure game Grad School: HGH is the final, extended version of a BME Prototyping class project in which the goal was to produce a zombie-themed game that teaches biomedical engineering concepts. The gameplay provides fast paced, exciting, and mildly addicting rooms that the player must battle and survive through, followed by an engineering puzzle that must be solved in order to advance to the next room. The objective of this project was to introduce the core concepts of BME to prospective students, rather than attempt to teach an entire BME curriculum. Based on user testing at various phases in the project, we concluded that the gameplay was engaging enough to keep most users' interest through the educational puzzles, and the potential for expanding this project to reach an even greater audience is vast.
ContributorsNitescu, George (Co-author) / Medawar, Alexandre (Co-author) / Spano, Mark (Thesis director) / LaBelle, Jeffrey (Committee member) / Guiang, Kristoffer (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor)
Created2014-05
Description
The American Diabetes Association reports that diabetes costs $322 billion annually and affects 29.1 million Americans. The high out-of-pocket cost of managing diabetes can lead to noncompliance causing serious and expensive complications. There is a large market potential for a more cost-effective alternative to the current market standard of screen-printed self-monitoring blood glucose (SMBG) strips. Additive manufacturing, specifically 3D printing, is a developing field that is growing in popularity and functionality. 3D printers are now being used in a variety of applications from consumer goods to medical devices. Healthcare delivery will change as the availability of 3D printers expands into patient homes, which will create alternative and more cost-effective methods of monitoring and managing diseases, such as diabetes. 3D printing technology could transform this expensive industry. A 3D printed sensor was designed to have similar dimensions and features to the SMBG strips to comply with current manufacturing standards. To make the sensor electrically active, various conductive filaments were tested and the conductive graphene filament was determined to be the best material for the sensor. Experiments were conducted to determine the optimal print settings for printing this filament onto a mylar substrate, the industry standard. The reagents used include a mixture of a ferricyanide redox mediator and flavin adenine dinucleotide dependent glucose dehydrogenase. With these materials, each sensor only costs $0.40 to print and use. Before testing the 3D printed sensor, a suitable design, voltage range, and redox probe concentration were determined. Experiments demonstrated that this novel 3D printed sensor can accurately correlate current output to glucose concentration. It was verified that the sensor can accurately detect glucose levels from 25 mg/dL to 400 mg/dL, with an R2 correlation value as high as 0.97, which was critical as it covered hypoglycemic to hyperglycemic levels. This demonstrated that a 3D-printed sensor was created that had characteristics that are suitable for clinical use. This will allow diabetics to print their own test strips at home at a much lower cost compared to SMBG strips, which will reduce noncompliance due to the high cost of testing. In the future, this technology could be applied to additional biomarkers to measure and monitor other diseases.
ContributorsAdams, Anngela (Author) / LaBelle, Jeffrey (Thesis advisor) / Pizziconi, Vincent (Committee member) / Abbas, James (Committee member) / Arizona State University (Publisher)
Created2017
Description
Over the past fifty years, the development of sensors for biological applications has increased dramatically. This rapid growth can be attributed in part to the reduction in feature size, which the electronics industry has pioneered over the same period. The decrease in feature size has led to the production of microscale sensors that are used for sensing applications, ranging from whole-body monitoring down to molecular sensing. Unfortunately, sensors are often developed without regard to how they will be integrated into biological systems. The complexities of integration are underappreciated. Integration involves more than simply making electrical connections. Interfacing microscale sensors with biological environments requires numerous considerations with respect to the creation of compatible packaging, the management of biological reagents, and the act of combining technologies with different dimensions and material properties. Recent advances in microfluidics, especially the proliferation of soft lithography manufacturing methods, have established the groundwork for creating systems that may solve many of the problems inherent to sensor-fluidic interaction. The adaptation of microelectronics manufacturing methods, such as Complementary Metal-Oxide-Semiconductor (CMOS) and Microelectromechanical Systems (MEMS) processes, allows the creation of a complete biological sensing system with integrated sensors and readout circuits. Combining these technologies is an obstacle to forming complete sensor systems. This dissertation presents new approaches for the design, fabrication, and integration of microscale sensors and microelectronics with microfluidics. The work addresses specific challenges, such as combining commercial manufacturing processes into biological systems and developing microscale sensors in these processes. This work is exemplified through a feedback-controlled microfluidic pH system to demonstrate the integration capabilities of microscale sensors for autonomous microenvironment control.
ContributorsWelch, David (Author) / Blain Christen, Jennifer (Thesis advisor) / Muthuswamy, Jitendran (Committee member) / Frakes, David (Committee member) / LaBelle, Jeffrey (Committee member) / Goryll, Michael (Committee member) / Arizona State University (Publisher)
Created2012