Functional 3-D Cellulose and Nitrocellulose Paper-based, Microfluidic Device Utilizing ELISA Technology for the Detection/Distinction Between Hemorrhagic and Ischemic Strokes

Abstract

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 a reasonable rate.For the biochemistry portion of this thesis project, antibodies were selected to target the intended stroke biomarker: GFAP. However, due to antibody pairing complications, the protein chosen for this project was natural human cardiac troponin T, which is elevated in the bloodstream of patients who have suffered a stroke. Several antibody experiments were performed to help finalize the procedure for performing an ELISA on the paper chip. The final antibody experiment was able to demonstrate that a paper microfluidic device utilizing ELISA techniques can successfully detect a stroke biomarker at physiologically relevant concentrations.Overall, this project supported the ability to accurately and effectively diagnose stroke in a timely manner through the use of a paper microfluidic device. v ACKNOWLEDGMENTS First and foremost I would like to thank my thesis advisor, Dr. David Clague. He was very supportive throughout the entire process and helped make key decisions in our brainstorming sessions. He was a knowledgeable advisor who was able to offer much guidance in this project, especially with the microfluidic concepts explored. He was very passionate about this project and was always hopeful even when we ran into issues; he would quickly make the changes needed to allow for the project to continue. I believe his passion for stroke research helped me realize the impact this project could have on thousands of people who have suffered a stroke.Secondly, I would like to thank my biology advisor, Dr. Amy Howes. She was an amazing mentor for the biology portion of the project and was so wonderful to work with. She