Bioimaging and Biomedical Optics Commons^™

Utilizing Spatial Transcriptomics To Compare Gene Expression In Volumetric Muscle Loss Injury Recovery, Colton Gattis

Biomedical Engineering Undergraduate Honors Theses

Volumetric Muscle Loss (VML) injuries are known to disrupt the normal regenerative process via fibrosis leading to an extensive permanent loss of muscle function. The specific causation of this disruption remains to be defined; however, with new developments in transcriptomics, genetic trends can be identified across regions of tissue over time. This study follows VML injuries within the tibialis anterior of a mouse model for fifteen days after initial injury to attempt to identify expected transitions in healing. Genetic presence began to become more diversified as recovery progressed from four to fifteen days post injury with spatial clusters becoming globalized …

Correlation Of Intracellular Metabolites With Optical Metabolic Imaging In Polarized Macrophages As Indicators Of Metabolic Reprogramming, Abby Claire Denison, Shelby Bess

Biomedical Engineering Undergraduate Honors Theses

Macrophages are a type of immune cell that are characterized by their ability to differentiate into various active phenotypes based on environmental stimuli. They can generally be classified as one of two extremes: inflammatory/M1 or wound-healing/M2. Macrophage populations in vivo, however, are rarely homogenous, and since macrophages play vital roles in the immune response, particularly in the context of the tumor microenvironment (TME), it’s important to be able to investigate and understand heterogenous macrophage populations to develop more effective treatments. Recently, autofluorescence imaging such as multiphoton microscopy and fluorescence lifetime imaging microscopy (FLIM) of metabolic cofactors NADH and FAD have …

Automated Segmentation Of Autofluorescence Lifetime Images Using A Convolutional Neural Network, Nicole Hooten

Biomedical Engineering Undergraduate Honors Theses

Chronic wounds affect nearly 1 out of every 50 people in the United States, decreasing quality of life and putting people with potential comorbidities at high risk for obtaining an infection. Wound healing progress in clinical settings is measured by tracking wound size and there are currently no non-invasive, quantitative measurement techniques. Previous studies have proposed nicotinamide adenine dinucleotide (NADH) autofluorescence lifetime (FLIM) imaging of the wound edge as a method to quantify the wound healing process by connecting the role of NADH in cellular metabolism to wound healing stage. However, evaluation of FLIM images is heavily subjective, as a …

Surface Laser Texturing Of Cocrmo Alloys, Gabriel Dharwadker

Biomedical Engineering Undergraduate Honors Theses

Artificial orthopedic implants such as hip joints suffer from excessive wear due to excess friction between the polyethylene and cobalt-chromium-molybdenum surfaces of the device. This increase in friction causes a fraction of them to give out within ten years. Therefore, to minimize the probability of this happening, recent research has been conducted regarding laser surface texturing of the cobalt-chromium-molybdenum alloy components of artificial joints. These surface textures serve to mimic the native lubrication mechanisms of cartilage in natural joints, which, in a natural joint, lowers the friction. However, the production of these laser surface textures is still a field of …

Assessing The Potential Effectiveness Of Diffuse Reflectance Spectroscopy For Quantifying Neonatal Skin Properties Using Tissue Phantoms, Jack Murphy

Biomedical Engineering Undergraduate Honors Theses

Diffuse reflectance spectroscopy (DRS) has previously been shown to be effective at analyzing the optical characteristics of a wide variety of biological tissues, including the skin. In addition to being sensitive to the amount of blood and its oxygenation, DRS can be used to extract bilirubin levels and assess jaundice, a common complication in newborn infants. To assess its potential for being utilized in applications involving neonatal skin, we evaluated the utility of DRS for extracting hemoglobin and bilirubin concentrations from tissue phantoms. Phantom sets were created by including a known amount of scattering and by varying known concentrations of …

Sequential Illumination In A Tomographic Microendoscopic Probe For Imaging Tumor Microvasculature, Zachariah Neumeier

Biomedical Engineering Undergraduate Honors Theses

Knowledge of colorectal cancer biology is improving how we approach cancer treatment. Specifically, the tumor microenvironment and abnormal angiogenesis are of particular interest. Optical methods are a prime candidate for research of the tumor microenvironment due to their ability to quantitively assess tissue structure and perfusion in real time. Particularly, the “transport scattering regime” has been identified as a method of obtaining high-resolution images and reflectance spectroscopy data; this light scattering regime has been demonstrated compatible with endoscopic imaging systems. In this study, a proof-of-concept optical imaging system is presented, capable of resolving absorbers within scattering turbid media using a …

The Effects Of Microgravity On Human Endothelial Cells In Relation To Deep Vein Thrombosis, Allie Burgess

Biomedical Engineering Undergraduate Honors Theses

Deep vein thrombosis (DVT) is a condition where blood clots form in one or more of the deep veins of the body, causing leg pain and/or swelling, potentially leading to pulmonary embolism and/or stroke. The incidence of DVT among astronauts serving on the International Space Station has been shown to be greater than that of the general population, calling into question the effects of microgravity on human thrombolytic processes. Additionally, microgravity conditions have been shown to cause muscular atrophy and a decrease in muscle strength and mobility. Because collagen composes a large portion of muscle tissue, this study also assessed …

Determination Of Tissue-Level Changes In Tumors That Are Indicative Of Metastasis Using Optical Spectroscopy, Sanidhya D. Tripathi

Biomedical Engineering Undergraduate Honors Theses

Breast cancer, accounting for 12% of all new cancer cases, is one of the leading causes of death in women worldwide. Although patient survival has improved over the years, metastatic spread to other organ sites and not due to the primary tumor is the most common form of tumor recurrence, accounting for 90% deaths. Hypoxia is a common hallmark of solid tumors and is linked with metastasis, therapeutic resistance, and poor patient survival. Defined as a state of decreased oxygen availability, cells under hypoxia have an increased rate of genetic mutation, local invasion, and resistance to treatment such as radiation …

3d Volumetric Mapping Of Tissue Properties Via A Catheter-Scale Microendoscope Imaging System, Andrew Stark

Biomedical Engineering Undergraduate Honors Theses

The study of tumor microvasculature is an important area of interest for research and clinical communities; however, there are significant limitations in the clinical characterization of the tumor microenvironment. Optical methods offer quantitative real-time measurement of tissue structure and perfusion and can be miniaturized for deployment endoscopically into previously inaccessible locations. However, conventional optical methods (i.e., optical coherence tomography, white light endoscopy, etc.) gives limited information about tissue perfusion while the diameter of various optical probes along with the complexity of scanning mechanisms make it difficult to access certain areas. In this study, a new technique for characterizing the tumor …

Investigating The Impact Of Hypoxia On Reactive Oxygen Species Generation Within Murine Breast Cancer Cells, Jared Mcpeake

Biomedical Engineering Undergraduate Honors Theses

When cancer metastasizes from a primary tumor site to secondary site through the bloodstream or lymph, the cancer becomes more difficult to treat. For this reason, it is vital to study what indicates the metastatic potential of a tumor. Current research has shown that cell lines with high metastatic potential display increased levels of metabolic adaptability over their nonmetastatic counterparts after undergoing hypoxic conditions. One method of assessing this adaptability is to measure the concentration of reactive oxygen species (ROS) produced by the cells while undergoing oxidative stress. In highly adaptable metastatic cells, an increase of ROS buildup within the …

Erythrocyte Deformability In Response To Glucose Using Liquid Crystals, Jayden Goff

Biomedical Engineering Undergraduate Honors Theses

The worldwide prevalence of diabetes mellitus is rapidly increasing with about 9.3% of the adult population living with the disease. People with diabetes have trouble regulating their blood glucose levels which typically leads to hyperglycemia. Under normal physiological conditions, erythrocytes can undergo deformations in response to shear stress when passing through capillaries with a smaller diameter. Poorly managed hyperglycemia can lead to the glycosylation of erythrocyte membrane proteins and hemoglobin. This glycosylation leads to increased rigidity of the cells along with decreased deformability in response to mechanical stress; therefore, these cells have a higher susceptibility of getting stuck in the …

Clot Analogs For The Development Of Improved Treatment Methods In Ischemic Stroke, Charles A. Rieth

Biomedical Engineering Undergraduate Honors Theses

According to the CDC, nearly 800,000 individuals experience stroke each year in the United States [1]. Greater than 70% of strokes are of ischemic etiology and involve the occlusion of key arteries in the cardiovascular system [2]. Tissue plasminogen activator (IV-tPA) is the current gold standard for thrombolytic approaches [3]; however, this therapeutic is only effective between 3 and 4.5 hours from the patient’s last know well [4]. As a result, less than 5% of acute ischemic stroke patients receive IV-tPA. In order to increase the viability of AIS treatment as a whole, it is essential that thrombolytic techniques are …

The Influence Of Mmp14 On Angiogenesis In Chemotherapy-Treated Tumors, Abdussaboor Muhammad

Biomedical Engineering Undergraduate Honors Theses

Colorectal cancer is the third most common cancer in the world, and it is the fourth most common cause of cancer related death (1). There have been many significant advancements regarding the treatment of cancer which aim to shrink the size of tumors in patients. However, there is still more that needs to be understood about the many different factors that play a role in colorectal cancer development.

Angiogenesis is the process of forming new blood vessels from existing ones and it requires breaking down and remodeling of the extracellular matrix (ECM) in order to allow endothelial cells to migrate …

Metabolic Profiling Of Radiation Resistance In Head And Neck Squamous Cell Carcinoma, Natalie Curry

Biomedical Engineering Undergraduate Honors Theses

Abstract

Head and neck squamous cell carcinoma is the sixth most common cancer worldwide. Radiation therapy is one of the most common treatments for HNSCC, however, many patients do not respond to radiation therapy. Currently, it takes about a month for clinicians to definitively determine whether a tumor is responsive to radiation therapy. Current studies have identified that the prevention of mitochondrial oxidative stress caused by reactive oxygen species can prevent DNA damage and lead to a decrease in apoptosis in radiation resistant cells Measuring the oxygen consumption rate through Seahorse metabolic assays allows us to quantify basal respiration, maximal …

Quantifying Blood Flow Patterns In The Pediatric Heart Using Blood Speckle Imaging, Harrison Dean

Biomedical Engineering Undergraduate Honors Theses

Congenital Heart Disease (CHD) is the most common type of congenital disease worldwide. Echocardiography using Doppler ultrasound is typically used to diagnose and monitor CHD; however, it is angle-dependent in nature and as a result is limited in its ability to accurately evaluate ventricular function. Blood Speckle Imaging (BSI) is a novel, angle-independent imaging modality that provides detailed blood flow information and visualization within a given area using speckle-tracking. In this study, velocity-field information obtained from BSI was used to calculate flow parameters such as kinetic energy (KE) and circulation using a custom created MATLAB program. BSI was performed on …

Quantifying Mucosal Hemodynamics In A Murine Model Of Ulcerative Colitis With Diffuse Reflectance Spectroscopy, Elizabeth A. Bullard, Ariel I. Mundo, Shelby N. Bess, Kathryn P. Miller

Biomedical Engineering Undergraduate Honors Theses

Ulcerative colitis (UC) is a gastrointestinal, autoimmune disease that causes ulceration and inflammation of the colon with an incidence 10 out of every 100,000 people in North America and Western Europe. Though the exact etiology is uncertain, a number of studies have shown that inflammatory cells along with environmental factors, genetics, and lifestyle habits can contribute to the sustained inflammatory response. In order to determine the cellular mechanism behind relapse and remission of UC, researchers have frequently employed immunohistochemistry, western blotting and gene sequencing, but these destructive analysis methods require the removal of a sample, necessarily limiting these methods to …

Quantification Of Superoxide Production In Cells Treated With L-Buthionine Sulfoximine, Rebekah Townsley

Biomedical Engineering Undergraduate Honors Theses

Hydroxyl radicals, superoxides, and hydrogen peroxides are common reactive oxygen species (ROS) that are produced by the mitochondria during cellular respiration [1]. Signal transduction pathways in the mitochondria are used to maintain the balance of ROS. However, when these mechanisms fail to regulate ROS properly, it causes oxidative damage and can lead to the development of various diseases such as cancer, respiratory, neurodegenerative, and cardiovascular diseases. L-buthionine sulfoximine (BSO) has been used previously to induce oxidative stress by inhibiting the γ-GCS enzyme used to synthesize glutathione, an antioxidant used to convert ROS to water [3]. BSO reduces glutathione production, thus …

Monitoring Fgf1-Treated Skin Wounds With Label-Free Multiphoton Microscopy, Gianna Busch

Biomedical Engineering Undergraduate Honors Theses

Chronic skin wounds pose a significant threat to public health, affecting as many as 5.5 million people in the United States every year and costing the healthcare system $10 billion annually^1,2. These wounds are associated with prolonged inflammation, poor vascularization, increased infection risk, and high mortality rates^3,4. There is significant interest in developing and testing a variety of biologics to promote wound healing. However, quantitative evaluations of healing are typically limited to measurements of wound size, and there is a critical need to develop quantitative biomarkers sensitive to different aspects of the healing process. Label-free multiphoton …

Developing Aligned Nerve Scaffolds In A 3d Type-I Collagen Gel, Gabriel David

Biomedical Engineering Undergraduate Honors Theses

Despite significant progress in the field of peripheral nerve repair, clinical success is still limited, leaving millions to suffer from peripheral neuropathy with billions spent every year for treatment. Nerve repair methods that are capable of maximizing the regenerative properties of peripheral nerves are greatly desired in the field of medical science. This research aims to fill the gap between modern methods and the future of nerve repair by creating type-I collagen scaffolds with aligned degradation pores that will assist and nurture nerves growing through them. This is achieved by incorporating adipose stem cells into type-I collagen hydrogels and aligning …

Autofluorescence Spectral Unmixing For Quantitative Metabolic Imaging Within Tissues, Lucy Woodbury

Biomedical Engineering Undergraduate Honors Theses

Label-free metabolic imaging through quantification of NADH and FAD autofluorescence has become a powerful and efficient tool for non-invasive measurements of cell metabolism. It has applications in a variety of fields including diagnostic and therapeutic monitoring. However, NADH and FAD imaging typically selects specific single excitation and emission bands where it is assumed that these fluorophores are individually isolated. In some cases of isolated cell imaging, this assumption holds true, but for samples with other intrinsic fluorophores present (i.e. collagen and elastin), it can interfere with quantitative results. Elastin and collagen autofluorescence prevent the broader application of these optical metabolic …

The Utilization Of Autofluorescence To Study The Effects Of L-Buthionine-Sulfoximine On Cellular Metabolism In Vitro, Madison Belew

Biomedical Engineering Undergraduate Honors Theses

Oxidative damage and stress impacts metabolic activity and has been implicated in a number of diseases . These medical conditions have been found to be associated with a lack of glutathione (GSH). There is a need to study the effects that the absence of glutathione has in order to expand knowledge of indicators that cause conditions like cataracts, cancer, or impaired wound healing. L-buthionine-sulfoximine (BSO) has been used in past studies to hinder GSH production and induce oxidative stress within a cell. This study explores the effects of BSO and induced oxidative stress on the metabolic activity and processes within …

Optical Spectrsocopy Of Murine Breast Tumor To Distinguish Indolent From Aggressive Disease, Joel Rodriguez Troncoso

Biomedical Engineering Undergraduate Honors Theses

Breast cancer accounts for 30% of all cancer. Metastasis is the primary cause of death among breast cancer patients. Additionally, current molecular profiling methods such as Oncotype DX, which are expensive and not widely available at all clinical facilities, only determine the risk of recurrence after treatment. Therefore, there are no current method capable of identifying metastatic patients in advance.As a result, there is an unmet clinical need to develop a cost-effective prognostic to differentiate between indolent and aggressive breast tumors. In this study, we implemented diffuse reflectance spectroscopy (DRS) system to evaluate functional changes in tumor xenografts originated from …

Vector Flow Imaging In Pediatric Cardiology - Extracting And Validating Data, Mason Belue

Biomedical Engineering Undergraduate Honors Theses

In the field of bedside cardiac diagnostic imaging, Doppler Ultrasound (DU) is the gold standard for diagnosing heart conditions. The largest benefit of DU is its ability to noninvasively image cardiac flow and allow the estimation of blood velocity and quantification of anatomical disease. However, to get correct velocity estimation, the position of the transducer in relation to the flow field needs to be known. This is the problem of angle/direction dependency and limits DUs accuracy when imaging in areas where perfect alignment or exact position of the transducer in relation to flow field is not possible or known, such …

Investigation Of Acute Radiation-Induced Changes In Oxygenation In A Murine Breast Tumor Model, Alaa Abdelgawad

Biomedical Engineering Undergraduate Honors Theses

Around 50-60% of all cancer patients undergo radiation therapy. Although some patients show complete response with no recurrence, a significant proportion of the population still develop radiation resistance. It is important to identify tumor resistance at early stages of therapy in order to adjust treatment protocol and avoid extra exposure to radiation. Current methods to assess treatment response are only limited to anatomical measurements of tumor volume after therapy. Novel approaches that shed the light on any functional information during the course of radiotherapy could significantly improve our ability to identify patients who do not respond to radiation therapy. Diffuse …

Modeling And Validation Of Tissue Optical Properties In The Photon Transport Regime, Katelyn Heath

Biomedical Engineering Undergraduate Honors Theses

Early detection of changes in epithelial cells, such as the development of neoplastic formations seen in epithelial dysplasia, can indicate regions of the epithelial tissue that are at a high risk for cancerous formation. Using concepts from diffuse reflectance spectroscopy, a Monte Carlo model was developed to predict the reflectance measured by a detector at a small source-detector separation on a microendoscope. The Monte Carlo results were then used to calculate a mathematical relationship between the reflectance and distance that can be used to determine optical properties in a tissue sample. This model was validated with liquid phantoms of specified …

Extraction And Analysis Of Vector Flow Imaging Data In A Pediatric Population, Bailey Stinnett

Biomedical Engineering Undergraduate Honors Theses

Vector flow imaging (VFI) is a new ultrasound technology that provides real time, angle-independent visualization of flow velocities in the heart and great vessels. Thus far, VFI has been used for superficial applications due to the limited penetration depth of available transducer probes; however, this depth in smaller pediatric patients enables adequate aortic views. In this project, VFI was used to study pediatric aortic stenosis (PAS)—a congenital heart defect that results in the narrowing of the aorta and/or aortic valve. The decision to refer PAS patients for surgical or catheter-based intervention is initially based on Doppler ultrasound. VFI is potentially …

Quantification Of Tumor-Associated Macrophages Following Immunomodulation Therapy In A Murine Allograft Model Of Colorectal Carcinoma, Caroline Spainhour

Biomedical Engineering Undergraduate Honors Theses

Colorectal Carcinoma (CRC) is one of the deadliest cancers in the world, with 150,000 new cases annually in the United States. Traditional treatments include chemotherapy and invasive surgery; however, research has shown that only 25% of patients that undergo traditional treatment have a positive result. Immunotherapy is an emerging form of cancer treatment that utilizes the patients’immune system to fight cancer cells by targeting inflammation, which plays a large role in the proliferation and metastasis of cancer cells.

Tumor-associated macrophages (TAMs) are immune cells that affect the inflammatory microenvironment of tumors. TAMs are M1 in the early stages of tumors, …

Comparison Of Varying Tissue Freezing Methods On Murine Colonic Tissue, James Hughes

Biomedical Engineering Undergraduate Honors Theses

Histology often requires a tissue specimen to be embedded so that it may be sectioned, stained, and mounted on a microscope slide for viewing. One common method of tissue embedding for rapid histology is freezing, since freezing allows tissue to be stored without the need for fixing. Frozen tissue is often embedded in a medium such as Optimal Cutting Temperature (OCT) compound so that it can be sectioned using a cryostat. However, factors such as ice-crystal formation during the freezing process can cause damage to the tissue. As such, the protocol used to freeze the tissue can affect the quality …

Metabolic Response To Stress Differentiates Heterogeneous Cancer Cells With Varying Metastatic Potential, Kinan Alhallak

Biomedical Engineering Undergraduate Honors Theses

Intratumoral heterogeneity is ubiquitously present within primary tumors and contributes to intractable behaviors such as metastasis and mutability spatiotemporally. Mounting evidence has shown that heterogeneous cell populations can adversely affect cell metabolism and metastatic potential. The cell’s only fluorescent molecules within the electron transport chain, flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide (NADH), can allow the quantitation of cell metabolism. We demonstrate the use of the optical redox ratio (FAD/(NADH+FAD)) to determine the metabolic behaviors of a heterogeneous panel of cells with varying metastatic programs at normal conditions and following acute hypoxia. At normal conditions, we reveal an attenuation …

Optical Imaging Of Metabolic Adaptability As A Biomarker For Metastatic Potential In Breast Cancer Cells, Mason G. Harper

Biomedical Engineering Undergraduate Honors Theses

Breast cancer metastasis is the main cause for mortality in breast cancer patients. However, knowledge of metastatic recurrence is limited, and there is a need to understand metastatic recurrence in order to treat breast cancer patients more effectively. Highly invasive metastatic breast cancer has shown to exhibit metabolic adaptability, transitioning from glycolysis to oxidative phosphorylation in the presence of microenvironmental stress. NADH and FAD are naturally occurring cofactor products during glycolysis and oxidative phosphorylation, respectively, and they are of particular importance during these metabolic processes due to their endogenous fluorescence. Measuring the ratio of fluorescence intensities of these cofactors through …