Bioimaging and Biomedical Optics Commons^™

Characterization Of The Response Of Trpv4 To Chemical Stimulation, Jacob V. Schluns

Biomedical Engineering Undergraduate Honors Theses

Traumatic Brain Injury (TBI) is a source of acute and chronic health issues for many patients. One of the components of the brain’s response to injury is astrogliosis, in which astrocytes that normally function to repair the brain instead form scar tissue that halts repair processes. Transient Receptor Potential Vanilloid Type 4 (TRPV4) is a trans-membrane calcium channel involved in astrogliosis. Through Fura-2AM based calcium imaging, the base activity of this channel in mouse astrocyte cells was recorded. The cells were then subjected to TRPV4 agonist and antagonist stimulation and their subsequent activity levels were recorded. The data showed that …

Optimizing A Standard Fasting Time For 2-Nbdg Uptake Studies In Murine Breast Cancers, Andrew C. Briley

Biomedical Engineering Undergraduate Honors Theses

Recently, there has been a larger use of 2-NBDG, a fluorescent glucose analog, to study glucose uptake in different cell types. These cell types have ranged anywhere from bacteria to human cancer cells. However, there has yet to be a standard procedure and practice for using 2-NBDG. In this study, our goal is to create a standard fasting time for the cells before introducing 2-NBDG to them. This study uses 4T07 cells, a murine breast cancer cell line, to help optimize this fasting time. These cells were fasted at different time points in order to find the ideal fasting time. …

In Vivo Multi-Parametric Imaging Of Metastatic And Non-Metastatic Breast Cancer, Raisa B. Rasul

Biomedical Engineering Undergraduate Honors Theses

A current issue in cancer therapy is the characterization of metastatic tumors, which can increase ease of treatment and patient trials. We present an in vivo study of metastatic (4T1) and non-metastatic (4T1-TWIST KO) breast tumor sister cell lines to understand their metabolic behavior, determine differences in two modes of imaging (reflection & transmission), and observe effect of breathing higher oxygen percentage on vascular hemoglobin oxygen saturation. After injection of 10,000 cells into mice dorsal window chambers, the glucose intake and hemoglobin oxygen saturation was measured using a fluorescent glucose analog (2-NBDG) and hyperspectral trans-illumination imaging from 520-620 nm at …

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 …

Determining The Effect Of Delivery Rate On Glucose Uptake By Cancer Cells, Luke Felton

Biomedical Engineering Undergraduate Honors Theses

Radiation therapy is one of the most commonly performed cancer treatment therapies today. Radiation therapy can cause can changes to tumor vasculature and affect blood flow rates. It is important to determine how changes in blood flow rates affect tumor glycolytic uptake to better understand tumor response to radiation therapy (1). The goal of this study was to use a fluorescent analog of glucose, 2-NBDG, to study the effect of varying flow rates on glucose uptake. A microfluidic device was constructed to flow 2-NBDG through. A murine metastatic breast cancer cell line was then cultured inside this microfluidic channel, and …

Optimized Protocol For Measuring 2-Nbdg Uptake As A Cellular Marker Of Glycolytic Demand, Drew Folgmann

Biomedical Engineering Undergraduate Honors Theses

2[N-(7-nitrobenz-2-oxa-1,2-diaxol-4-yl)amino]-2-deoxyglucose (2-NBDG) a fluorescently tagged analog of FDG is ideal for in vitro studies and imaging. 2-NBDG has been proven to be an adequate marker for glucose uptake in many different types of cells [1]. However, across the spectrum of 2-NBDG use a lack of consensus is observed for the following questions. What is the ideal time to fast cells to optimize cellular uptake of 2-NBDG? Also, what is the ideal concentration of 2-NBDG to be used when quantifying glucose uptake? Finally, what is the effect of serum on the uptake of 2-NBDG? To answer these questions and understand glucose …

Design Of A 3d-Printed Device For Diffuse Reflectance Spectroscopy Calibration As A Step Towards Clinical Translation, Nontapoth Vongkittiargorn

Biomedical Engineering Undergraduate Honors Theses

The diffusive reflectance and spectroscopic microendoscopy (DRSME) is a multimodal imaging system that harnesses its usefulness from different light sources. One of the modalities, diffuse reflectance spectroscopy (DRS), has been used in our lab to investigate optical properties of epithelial tissues using a broadband white light as the main source. Calibration of DRS is required as it can obtain the maximum intensity and convert it to absolute reflectance. Current manual-adjusted calibration can be lengthy and often lead to inconsistent results. Therefore, a new method of calibration is introduced where additive manufacturing (or 3D printing) technology is fully utilized.

The instrumentation …

Multiphoton Imaging Of Labeled Breast Cancer Cells To Quantify Intra And Extracellular Receptors, Sydney C. Wiggins

Biomedical Engineering Undergraduate Honors Theses

Every year 200,000 women in the United States are diagnosed with breast cancer. Of the cases diagnosed, 10% -15% are classified as triple negative breast cancer (TNBC) due to the absence of estrogen, progesterone, and HER-2/Neu receptors. This breast cancer sub-type is markedly more aggressive and twice as likely to develop in premenopausal women. TNBC is resistant to endocrine therapies and current targeted agents, making clinical need for the development of validated therapeutics for TNBC a pressing matter. To initiate drug development, the internalization of directly immunolabeled epidermal growth factor receptors (EGFR) in SK-BR-3 human breast adenocarcinoma cells was quantitated …

Intravital Microscopy Of Tumor Oxygenation And Glycolytic Demand, Jesse D. Ivers

Biomedical Engineering Undergraduate Honors Theses

There is growing concern about the over treatment of cancer because treatments are based primarily on tumor anatomy. In this study, we aim to begin the process of addressing that issue by developing an intravital technique for optically analyzing tumor biology. Two traits were identified as having significant importance in the aggression of a tumor, vascular oxygenation (SO2) and glycolytic demand. Dorsal skin flap window chambers were implanted and 4T1 and 67NR cancer cells were injected to provide a tumor model for the development of this intravital quantification technique. This study provides a detailed protocol from instrumentation setup to surgical …