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An Automated Lab-On-A-Cd System for Parallel Whole Blood Analyses

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An Automated Lab-On-A-Cd System for Parallel Whole Blood Analyses AN AUTOMATED LAB-ON-A-CD SYSTEM FOR PARALLEL WHOLE BLOOD ANALYSES (Spine title: A Lab-on-a-CD System for Blood Analyses) (Thesis format: Integrated Article) by Tingjie Li Department of Mechanical and Materials Engineering Faculty of Engineering A thesis submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy The School of Graduate and Postdoctoral Studies The University of Western Ontario London, Ontario, Canada © Tingjie Li 2012 THE UNIVERSITY OF WESTERN ONTARIO School of Graduate and Postdoctoral Studies CERTIFICATE OF EXAMINATION Supervisor Examiners ______________________________ ______________________________ Dr. Jun Yang Dr. Andy (Xueliang) Sun Supervisory Committee ______________________________ Dr. Christopher G. Ellis ______________________________ Dr. Andy (Xueliang) Sun ______________________________ Dr. George K Knopf ______________________________ Dr. Liying Jiang ______________________________ Dr. Xinyu Liu The thesis by Tingjie Li entitled: An Automated Lab-on-a-CD System for Parallel Whole Blood Analyses is accepted in partial fulfillment of the requirements for the degree of Doctor of Philosophy ______________________ _______________________________ Date Chair of the Thesis Examination Board ii Abstract Medical diagnostics plays a critical role in human healthcare. Blood analysis is one of the most common clinical diagnostic assays. Biomedical engineers have been developing portable and inexpensive diagnostic tools that enable fast and accurate tests for individuals who have limited resources in places that require such field applications. The emergence of Lab-on-a-CD technology provides a compact centrifugal platform for high throughput blood analysis in point-of-care (POC) diagnostics. The objective of this thesis work is to develop a Lab-on-a-CD system for parallel quantitative detection of blood contents. Blood separation is a key step in blood analysis. By integrating out-of-plane microvalves into the Lab-on-a-CD system, pure plasma is separated from human whole blood sample for subsequent blood detection. These out-of-plane microvalves show excellent performance in preventing backflow or reverse mixing due to blood cell diffusion. The concentrations of certain blood contents in the separated plasma can be detected using the electrochemical biosensors embedded in the plasma reservoir. To improve sensitivity, a nanoporous structure is created on the surfaces of the biosensors’ electrodes through an alloying/dealloying process. The nanoporous electrode has an electroactive surface area up to 60 times larger than that of a flat gold electrode of the same size. As a result, numerous catalysts and enzymes are stably entrapped in the nanoporous structure, leading to high sensitivity, stability and reproducibility of the biosensor. Based on this devised blood separation technique and the improved electrochemical detection method, a Lab-on-a-CD prototype was constructed and successfully applied in the concentration detection of glucose, lactate and uric acid with linear ranges of 0 – 30 mM, 0 – 1.5 mM and 0 – 5 mM, respectively. Furthermore, the volume of whole blood sample consumed for each section can be as small as 16 µL. iii The Lab-on-a-CD platform developed in this thesis is low-cost, robust, and simple-to-use. Potentially, it could be used in clinical diagnostics and will especially aid developing countries where resources are limited. Keywords Lab-on-a-CD, point-of-care, blood analyses, out-of-plane microvalves, electrochemical detection. iv Co-Authorship Statement This doctoral thesis has been carefully prepared according to the regulations for an integrated-article format thesis stipulated by the Faculty of Graduate and Postdoctoral Studies at the University of Western Ontario, and has been co-authored as follows: CHAPTER 3: OUT-OF-PLANE MICROVALVES FOR WHOLE BLOOD SEPARATION ON LAB-ON-A-CD All the theoretical analyses were conducted by T. Li under the supervision of Dr. J. Yang. Device fabrication and experimental testing were undertaken by T. Li under the supervision of Dr. J. Yang. Drafts of Chapter 3 were prepared by T. Li and reviewed by Dr. J. Yang. A paper co-authored by T. Li, L. Zhang and J. Yang has been published on Journal of Micromechanics and Microengineering. CHAPTER 4: DESIGN AND OPTIMIZATION OF PRUSSIAN BLUE/CARBON NANOTUBE ELECTRODES FOR BIOSENSING APPLICATIONS All the theoretical analyses were conducted by T. Li under the supervision of Dr. J. Yang. Material synthesis and experimental testing were undertaken by T. Li under the supervision of Dr. J. Yang. Drafts of Chapter 4 were prepared by T. Li and reviewed by Dr. J. Yang. A paper co-authored by T. Li, J. Yang, Q. Zhang has been submitted to Analytical Letters. CHAPTER 5: FABRICATION OF NANOPOROUS THIN-FILM WORKING ELECTRODES AND THEIR BIOSENSING APPLICATIONS All the theoretical analyses were conducted by T. Li under the supervision of Dr. J. Yang. Material synthesis and experimental testing were undertaken by T. Li under the supervision of Dr. J. Yang. Drafts of Chapter 5 were prepared by T. Li and reviewed by Dr. J. Yang. A v paper co-authored by T. Li, F. Jia, Y. Fan, Z. Ding and J. Yang has been submitted to Biosensors and Bioelectronics. CHAPTER 6: A ROBUST LAB-ON-A-CD SYSTEM FOR HIGH-THROUGHPUT AND AUTOMATED WHOLE BLOOD ANALYSIS All the theoretical analyses were conducted by T. Li under the supervision of Dr. J. Yang. Device fabrication and experimental testing were undertaken by T. Li under the supervision of Dr. J. Yang. Drafts of Chapter 6 were prepared by T. Li and reviewed by Dr. J. Yang. A paper co-authored by T. Li and J. Yang is to be submitted. vi Acknowledgments After been through four years hard work, I am completing my PhD degree in the summer of 2012. It has been my honor to spend these years in the Department of Mechanical and Materials Engineering at the University of Western Ontario, and its faculty and staff members will always remain dear to me. The best and worst moments of my doctoral journey have been shared with many people. My deepest gratitude must go to my supervisor, Dr. Jun Yang, for introducing me to this state-of-the-art research area and for his continuous guidance and motivation to me to think outside the box. Besides thesis research, Dr. Yang also taught me how to write research proposals and got me involved an industry/university collaboration project. All of these help me become a qualified Ph.D. Members of the Lanxess project, including Dr. Leo Lau, Dr. Liying Jiang, Dr. Natalie Suhan, Mr. Lorenzo P. Ferrari, Dr. Gilles Arsenault also deserve my sincerest thanks, their assistance and friendship has meant more to me than I could ever express. Special thanks to technical support from Surface Science Western, especially Mr. Brad Kobe, Mr. Ross Davidson, Ms. Marry Jane Walzak, and Dr. Heng-yong Nie. I also appreciate the technical support from the staff at the Western Nanofabrication Facility, including Mr. Tim Goldhawk, Dr. Todd Simpson, and Dr. Rick Glew. I would also like to thank the thoughtful discussion and friendly support from my colleagues and friends. Finally, I wish to express my highest gratefulness to all my family members. Their love provided my inspiration and was my driving force. I owe them everything and wish I could show them just how much I love and appreciate them. vii Table of Contents CERTIFICATE OF EXAMINATION ........................................................................... ii Abstract .............................................................................................................................. iii Co-Authorship Statement.................................................................................................... v Table of Contents ............................................................................................................. viii List of Tables .................................................................................................................... xii List of Figures .................................................................................................................. xiii Nomenclature .................................................................................................................... xx Chapter 1 ............................................................................................................................. 1 1 Introduction ...................................................................................................................... 1 1.1 Background .............................................................................................................. 1 1.2 Research Objectives and Outline of this Thesis ....................................................... 2 1.3 References ................................................................................................................ 4 Chapter 2 ............................................................................................................................. 5 2 Literature Review ............................................................................................................. 5 2.1 Point-of-care Diagnostics ......................................................................................... 5 2.2 Lab-on-a-CD Technology ...................................................................................... 11 2.2.1 Microfluidic Pumps ..................................................................................... 12 2.2.2 Microfluidic
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