Biomedical Engineering and Bioengineering Commons^™

Primer Payload System For Higher-Order Multiplex Lamp: Design And Development Of Unit Processes, Tochukwu Dubem Anyaduba

KGI Theses and Dissertations

Design and Development of Platforms for the Application of Loop-mediated Isothermal Nucleic Acid Amplification, LAMP, in the Diagnosis of Polymicrobial Diseases

Tochukwu Dubem Anyaduba, Travis Schlappi (PI)

For the past two decades, several isothermal nucleic acid amplification technologies have emerged. These are mostly in response to the need for robust molecular diagnostic tools amenable to point-of-care and limited-resource settings. Of these, loop-mediated isothermal amplification, LAMP, stands out as a highly specific and rapid alternative to the polymerase chain reaction, PCR. One of LAMP's significant characteristics involves using four essential and two loop (rate increasing) primers to recognize six to eight …

Two-Dimensional & Three-Dimensional Microarray Cell Culture Using Elastomeric Assembly Substrates, Angel Olivera-Torres

Honors Program: Senior Projects (Public)

Tissue engineering and regenerative medicine represent the collection of all engineering disciplines brought together for the common goal of developing novel ways of growing tissues and organs in the laboratory. Efforts have made it possible to replicate or induce growth of 2D structures in the human body like skin, but the clinical need for on-demand solid organs has yet to be met due to lack of understanding of the variables responsible for organogenesis. Cell-colony heterogeneity, 3D-cellular architecture, bioactive molecules, and crosstalk communication between parenchymal cell populations need to be further investigated, and high-throughput technologies can rapidly increase the rate at …

Dynamic Behavior And Interactions Of Ferrofluid Droplets Under Magnetic Fields In Low Reynolds Number Flows, Md Rifat Hassan

Doctoral Dissertations

Digital microfluidics in combination with emulsion microfluidics are crucial building blocks of droplet-based microfluidics, which are prevalent in a wide variety of industrial and biomedical applications, including polymer processing, food production, drug delivery, inkjet printing, and cell-based assays. Therefore, understanding the dynamics and interactions of droplets as well as the interactions between the droplets and solid surfaces are of great importance in order to improve the performance or product in these applications.

Recently, several studies in the literature have demonstrated the potential of magnetic fields in controlling the behavior of droplets in microscale; however, the fundamental mechanism behind the interesting …

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 …

Bioprinted In Vitro Model Of Human Glioblastoma, Rachel Lauren Schwartz

Theses and Dissertations

Glioblastoma multiform (GBM) is one of the most aggressive forms of primary brain tumors. GBM is fast progressing and resistant to treatment, resulting in a low survival rate. Conventional 2-dimensional tissue culture models cannot fully replicate the complexities of cancer lesions that contain multiple cell types and structures (e.g. vessels composed of endothelial cells, cancer cells, normal cells, etc.) as well as an intricate scaffold of proteins comprising the extracellular matrix (ECM). In addition, animal models cannot translate into the clinical disease in patients. Thus, this study has developed a bioprintable organ-on-a-chip (OOAC) model that mimics the important ECM factors …

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 …

Modification And Optimization Of Conducting Polymer-Modified, Redox-Magnetohydrodynamics (R-Mhd) Pumping For Enhanced And Sustained Microfluidics Applications, Md Foysal Zahid Khan

Graduate Theses and Dissertations

In this work, a novel microfluidic pumping approach, redox-magnetohydrodynamics (R-MHD) has improved by materials and device optimization to use in lab-on-a-chip applications. In R-MHD, magnetic flux (B) and ionic current density (j) interacts to generate body force (FB) in between active electrodes, according to the equation FB = j×B. This unique fluid pumping approach is scalable, tunable, generates flat flow profile, and does not require any channels or valves. Pumping performance, such as speed scales with the ionic current density (j) and duration depends on the total charge (Q). The ionic current density (j) results from the conversion of electronic …

A Microfluidics-Based Cross-Flow Filtration Platform For Rapid Processing Of Amphiphilic Biomarkers From Blood, Kiersten D. Lenz

Biomedical Engineering ETDs

Early and accurate detection of bacterial infections can help save lives, prevent the spread of disease, and decrease the overuse of antibiotics. Our team at the Los Alamos National Laboratory has developed novel assays to detect bacterial biomarkers from patient blood at the point-of-care in order to facilitate a universal diagnostic platform. However, these biomarkers are amphiphilic in nature, and this biochemical property causes them to be sequestered by high-density and low-density lipoproteins (HDL and LDL) in the host’s blood. Extraction of the bacterial biomarkers from the lipoprotein complexes is thereby required for the development and deployment of a diagnostic …

Microfluidic Chip For High Efficiency Microinjection Of Caenorhabditis Elegans, Delaney Gray, Alex Hadsell, Jessica Talamantes

Bioengineering Senior Theses

The terrestrial nematode, Caenorhabditis elegans, is an invaluable model organism for the study of molecular and cellular processes due to their small size, rapid generation time, easy cultivation, and invariant cell number. Additionally, 40% of genes known to be associated with human disease have clear orthologs in the C. elegans genome. In C. elegans genetics research, microinjection of genetic material into the worms is critical. Although an established technique, manual microinjection is tedious, low-throughput, and requires an expert researcher. This thesis details a novel microfluidic device designed to perform high-throughput microinjection. This two-layer, PDMS-based chip integrates microfluidic elements to …

3d Printing A Microfluidic Chip Capable Of Droplet Emulsion Using Ninjaflex Filament, Robert Andrews

Mechanical Engineering Undergraduate Honors Theses

This paper details an investigation into methods and designs of 3D printing a microfluidic system capable of droplet emulsion using NinjaFlex filament. The specific field in which this paper’s experiment is rooted is dubbed “BioMEMS,” short for bio microelectromechanical systems. One prominent research area in BioMEMS is developing a “lab on a chip.” Essentially, the goal is to miniaturize common lab processes to the micro scale, rendering it possible to include these processes in a small chip. Reducing necessary sample sizes, shortening the reaction times of lab processes, and increasing mobility of lab processes can all be realized through microfluidic …

Use Of Surface-Enhanced Raman Scattering (Sers) Probes To Detect Fatty Acid Receptor Activity In A Microfluidic Device, Han Zhang, Wei Zhang, Lifu Xiao, Yan Liu, Timothy A. Gilbertson, Anhong Zhou

Biological Engineering Faculty Publications

In this study, 4-mercaptobenzoic acid (MBA)-Au nanorods conjugated with a GPR120 antibody were developed as a highly sensitive surface-enhanced Raman spectroscopy (SERS) probe, and were applied to detect the interaction of fatty acids (FA) and their cognate receptor, GPR120, on the surface of human embryonic kidney cells (HEK293-GPRR120) cultured in a polydimethylsiloxane (PDMS) microfluidic device. Importantly, the two dominant characteristic SERS peaks of the Raman reporter molecule MBA, 1078 cm⁻¹ and 1581 cm⁻¹, do not overlap with the main Raman peaks from the PDMS substrate when the appropriate spectral scanning range is selected, which effectively avoided the …

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 …

Progressive Hypoxia-On-A-Chip: An In Vitro Oxygen Gradient Model For Capturing The Effects Of Hypoxia On Primary Hepatocytes In Health And Disease, Young Bok Abraham Kang, Jinsu Eo, Beyza Bulutoglu, Martin L. Yarmush, O. Berk Usta

Faculty Publications - Biomedical, Mechanical, and Civil Engineering

Oxygen is vital to the function of all tissues including the liver and lack of oxygen, that is, hypoxia can result in both acute and chronic injuries to the liver in vivo and ex vivo. Furthermore, a permanent oxygen gradient is naturally present along the liver sinusoid, which plays a role in the metabolic zonation and the pathophysiology of liver diseases. Accordingly, here, we introduce an in vitro microfluidic platform capable of actively creating a series of oxygen concentrations on a single continuous microtissue, ranging from normoxia to severe hypoxia. This range approximately captures both the physiologically relevant oxygen gradient …

Neuron-Glial (Ng) Interactions: A Microfluidic Examination Of Ng Emergent Responses For Repair, Tanya Singh

Dissertations and Theses

Neuron-glia communication is crucial to the development, plasticity, and repair of the nervous system (NS). While neurons are well known to conduct electrical impulses that transfer biological information and stimuli throughout the NS, our understanding of the roles of glia continues to evolve from when the cells were largely believed to act solely for neuronal support. Recent decades of research has shown that glia can alter metabolism, conduct impulses and change phenotype for NS repair. NG interactions have, thereby, become heavily researched in varied areas of biomedical engineering, including embryogenesis, neural regeneration, growth, and intracellular synaptic activity. However, while NG …

Controlled Migration Of Retinal Progenitor Cells Within Electro-Chemotactic Fields, Shawn Mishra

Dissertations and Theses

Vision loss in retinal degenerative diseases is overwhelmingly attributed to damage and death of retinal photoreceptor cells. Studies in mouse retina have suggested that transplantation of isolated post-natal or stem cell-derived retinal progenitor cells (RPCs) to replace apoptotic or damaged photoreceptors may be a novel approach to restore vision. Thus far, outcomes project that the amount of restored visual response depends upon the migration of transplanted cells from insertion in the sub-retinal space to the outer nuclear layer (ONL). However, transplantation efficiency is exceedingly low – ~5% cells transplanted enter the retina – directly limiting the efficacy of the treatments. …

Collective Behavior Of Drosophila Melanogaster Neural Progenitor And Imaginal Disc Cells Within Controlled Microenvironments, Caroline D. Pena

Dissertations and Theses

Regenerative therapies for the damaged visual system have introduced stem-derived cells to recapitulate developmental processes and initiate functional regeneration in different components of the eye. The developing visual system in Drosophila Melanogaster offers a model in which to analyze the associated processes in retinogenesis. The optic nerve is critical to vision and is developmentally preceded in Drosophila by a structure called the Optic Stalk (OS). Collective migration of neural and retinal progenitor cells (RPCs) from the developing brain lobes (DBL) to the Imaginal Disc (ID), through the OS, is a fundamental part of regenerative strategies in retina. Developmental signals governing …

Acoustofluidics And Soft Materials Interfaces For Biomedical Applications, Frank A. Fencl

Biomedical Engineering ETDs

This dissertation describes fabrication of devices and other tools for biomedical applications through the integration of acoustofluidic systems with bio separation assays, instrumentation components, and soft materials interfaces. For example, we engineer a new class of transparent acoustic flow chambers ideal for optical interrogation. We demonstrate efficacy of these devices by enhancing the signal for high throughput acoustic flow cytometry, capable of robust particle focusing across multiple parallel flowing streams. We also investigate an automated sampling system to determine the parameters of transient particle stream focusing in between sample boluses and air bubbles to model a high throughput, multi-sampling acoustic …

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 …

Direct Quantification Of Deubiquitinating Enzyme Activity In Single Intact Cells, Nora Safabakhsh

LSU Doctoral Dissertations

Challenges in drug efficacy occur during the treatment of most types of cancer due to the heterogeneity of the tumor microenvironment. This has led to the development of personalized medicine. Due to the clinical success of the proteasome inhibitors Bortezomib and Carfilzomib in treatment of multiple myeloma, interest has shifted towards molecularly-targeted chemotherapeutics for ubiquitin-proteasome system (UPS). Deubiquitinating enzymes (DUBs) are an essential part of this pathway which have been found to promote Bortezomib resistance in multiple myeloma patients. Unfortunately, there is a lack of specific, high throughput biochemical assays to characterize DUB activity in patient samples before and after …

Developing Droplet Based 3d Cell Culture Methods To Enable Investigations Of The Chemical Tumor Microenvironment, Jacqueline Alexandria De Lora

Biomedical Sciences ETDs

Adaptation of cancer cells to changes in the biochemical microenvironment in an expanding tumor mass is a crucial aspect of malignant progression, tumor metabolism, and drug efficacy. In vitro, it is challenging to mimic the evolution of biochemical gradients and the cellular heterogeneity that characterizes cancer tissues found in vivo. It is well accepted that more realistic and controllable in vitro 3D model systems are required to improve the overall cancer research paradigm and thus improve on the translation of results, but multidisciplinary approaches are needed for these advances. This work develops such approaches and demonstrates that new droplet-based cell-encapsulation …

Milkguard: Low-Cost, Polymer-Based Sensor For The Detection Of Escherichia Coli In Donated Human Breast Milk, Maggie May, Nicholas Kikuchi, Matthew Zweber

Bioengineering Senior Theses

Breast milk, the gold standard for infant nutrition, could prevent up to 13% of child deaths worldwide. However, many mothers are unable to breastfeed due to health conditions and other factors. Because of this, a network of more than 500+ human milk banks, which collect and distribute donated breast milk to infants, have emerged worldwide. However, operational costs to ensure the safety of this milk remain time-intensive and costly.

There are no existing diagnostics for rapid and on-site detection of bacterial contaminants in donated milk. Currently, many milk banks send samples to outside laboratories for bacterial culturing tests, which take …

Tumor-Targeted Double Emulsions For Ultrasound-Triggered Delivery Of Molecular Therapeutics., Connor S Centner

Electronic Theses and Dissertations

Cancer is the second leading cause of death in the United States, with 1.74 million new cancer cases diagnosed and 610,000 cancer deaths expected in 2018 alone. Current treatments often result in negative systemic effects and ineffective treatment of the tumor. Drug delivery vehicles have been developed for more effective local delivery methods, but many drug delivery vehicles lack spatial and temporal control. Targeted double emulsions are a new class of drug delivery vehicles which present a promising option for a high payload and controlled delivery. The purpose of our project was to develop and characterize an aptamer-chelated double emulsion …

Microfluidic Technology And Application In Urinal Analysis, Jiwen Xiang

LSU Doctoral Dissertations

Microfluidic technology offers numerous advantages in minimizing and integrating the traditional assays. However, the lack of efficient control components of the microfluidic systems has been hindering the widely commercialization of the technology. The research work in this dissertation focused on the development of effective control components for microfluidic applications.

A linear peristaltic pump was firstly designed, fabricated, and tested for conventional microfluidics by synchronously compressing the microfluidic channel with a miniature cam-follower system in Chapter 2. The miniature cam-follower system and microfluidic chip was prototyped using three-dimensional (3D) printing technology and soft lithography technology. Results from experimental test showed that …

Developing A 3d In Vitro Model By Microfluidics, Hung-Ta Chien

Dissertations and Theses

In vitro tissue models play an important role in providing a platform that mimics the realistic tissue microenvironment for stimulating and characterizing the cellular behavior. In particular, the hydrogel-based 3D in vitro models allow the cells to grow and interact with their surroundings in all directions, thus better mimicking in vivo than their 2D counterparts. The objective of this thesis is to establish a 3D in vitro model that mimics the anatomical and functional complexity of the realistic cancer microenvironment for conveniently studying the transport coupling in porous tissue structures. We pack uniform-sized PEGDA-GelMA microgels in a microfluidic chip to …

Collective Chemotaxis Of Retinal Neural Cells From Drosophila Melanogaster In Controlled Microenvironments, Stephanie Zhang

Dissertations and Theses

More than 172 million people are influenced by a retinal disorder that stems from either age-related or developmental causes. Of those, 1.5 million people endure a developmental retinal disorder. In the developing retina, neural cells undergo a series of highly complicated differentiation and migration process. A main cause of these diseases is abnormal collective migration of neural progenitors hindering the retinogenesis process. However, our grasp of collective migration and signaling molecules, critical to the developing retina, is incompletely understood. Understanding the molecular mechanisms, such as the fibroblast growth factor pathway, that regulate glial and neuronal migration provides decisive insights in …

Three-Dimensional Microfluidic Tumor Vascular Model For Investigating Breast Cancer Metastasis, Anastasiia Vasiukhina, Brian H. Jun, Luis Solorio, Pavlos P. Vlachos

The Summer Undergraduate Research Fellowship (SURF) Symposium

Metastasis is one of the primary reasons for the high mortality rates in female patients diagnosed with breast cancer. It involves the migration of cancer cells into the circulatory system allowing for the dissemination of cancer cells in distal tissues. Understanding the major processes that occur in cells and tissues during metastasis can help improve currently existing therapeutic methods. In order to understand such mechanisms, developing physiologically relevant tissue models is crucial. Advancements in microfluidics have led to the fabrication of 3D culture models with shear stress gradients and flow control that can recapitulate aspects of the tumor microenvironment in …

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 …

Shear Driven Micro-Fluidic Pump For Cardiovascular Applications, Nihad E. Daidzic

Aviation Department Publications

A valveless shear-driven micro-fluidic pump design (SDMFP) for hemodynamic applications is presented in this work. One of the possible medical and biomedical applications is in-vivo hemodynamic (human blood circulation) support/assist. One or more SDMFPs can be inserted/implanted into vascular lumens in a form of a stent/duct in series and/or in parallel (bypass duct) to support blood circulation in-vivo. A comprehensive review of various micro-pump designs up to about mid 2000’s is given in [1,2]. Many of micropump designs considered are not suitable for in-vivo or even in-vitro medical/biomedical applications.

Operating principles, design, and SDMFP features are given in [3]. A …

Improvement Of Fluorescence-Based Microfluidic Dna Analyzers, Collin Tranter

Doctoral Dissertations

A tremendous effort continues in the development of micro-total-analysis-systems; in support of this, many chemical passivation methods have been developed to enhance the biocompatibility of such microfluidic systems. However, the suitability of these passivation techniques to many fluorescence-based assays still remains inconsistent. This part of this work is focused on the performance of a third generation intercalating DNA dye when used within microfluidic devices treated with a select variety of passivating coatings. The results of these tests indicate that passivation coatings which are intended to shed DNA based on electrostatic repulsion will in fact imbibe the fluorescent DNA intercalating dye …