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Real Time Measurement of Oxygen by Integrating a Clark Sensor with Low Cost Printed Circuit Board Technology and Solid Electrolyte Membrane

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Real Time Measurement of Oxygen by Integrating a Clark Sensor with Low Cost Printed Circuit Board Technology and Solid Electrolyte Membrane Real Time Measurement of Oxygen by Integrating a Clark Sensor with Low Cost Printed Circuit Board Technology and Solid Electrolyte Membrane By Ayda Niazi A thesis submitted to The University of Birmingham for the degree of DOCTOR OF PHILOSOPHY COLLEGE OF ENGINEERING AND PHYSICAL SCIENCE SCHOOL OF MECHANICAL ENGINEERING JANUARY 2016 University of Birmingham Research Archive e-theses repository This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder. Abstract A prototype of a miniaturized Clark type electrochemical oxygen sensor integrated with a 3D printed in vitro cell culturing platform is designed and developed for the purpose of monitoring the cellular oxygen consumption by the solution flowing through the cultured cells on the platform. Oxygen respiration indicates a cell's metabolic activity, so by measuring a chemical's oxygen content as it passes through a cell chamber, we can measure that chemical's potential effectiveness. This miniature micro sensor is designed and fabricated on a printed circuit board for the first time and integrated with a solid electrolyte membrane and 3D printed cell culturing platform to ensure robustness and low manufacturing cost. Hence the sensor is aimed at enabling the pharmaceutical industry to rapidly test chemical products on animal and cancer cells; and has been designed to be low cost and suitable for mass production. The cell culturing platform’s prototype, which contains inlet and outlet pipes and a cell culturing chamber, was manufactured by The Eden250™ 3D Printing System using the Objet biocompatible material MED610™. The presented oxygen sensor configuration consists of two identical series of working, reference and counter microelectrodes, placed before and after the cell culturing chamber. It was manufactured by combining printed circuit board technology and laser micro machining techniques to create microscale microelectrode spacing. The usual electrolyte solution is replaced with a solid polymer electrolyte membrane, Nafion (perfluorosulfunic acid membrane, DuPont Company), which removed requirement for extra humidification and increased the shelf life of the dissolved oxygen sensing system. The solid polymer electrolyte also ensured robustness and good electrical conductivity for sensing. The sensor functions without humidification or any special condition and has a long shelf life. The sensitivity of the oxygen sensor was tested in different oxygen concentration in gas and liquid states and was calibrated with measurements II from a Portable Multi-Gas Analyser provided by Super Systems Europe and a Gold probe dissolved oxygen analyser. The prototype can detect the small changes in oxygen concentration in the range of 0 to 5 μA current and has a response time of less than 5 seconds. III I dedicate this thesis to my parents. IV Acknowledgment First I would like to thank my supervisor Dr. Carl Anthony for his kind help, wisdom and supervision during the period of my research course. I wish to thank Super Systems Europe group for their productive cooperation and their kind help to provide sensor calibration testing facilities. I would like to thank my colleagues Aydin, Nasim, Tianshi, Ali Kubba, and Haseena at the micro and nano technology centre of Birmingham University and my dear friends Fatima and Mahyar. I also wish to thank department of Chemical Engineering to provide Gold probe dissolved oxygen analyser to calibrate my sensors. I would like to specially thank Mr. Alan Yates for his kind help and guidance with circuit design and fabrication. I would particularly like to thank my father, Annemohammed Niazi for his unwavering moral, emotional and financial support and endless inspiration and my mother, sister and brothers, Sarah, Anya, Nasin, Poulad, Behzad and Timor for their love and emotional support throughout my academic life. Finally I would like to thank my best friend James for his endless encouragement, love and support throughout the period of my research course. Ayda Niazi Birmingham January 31, 2016 V List of Publications During the course of this project, the following Conference Proceedings have been published: Ayda Niazi, Carl Anthony, Development of oxygen sensor by integrating the low cost printed circuit board technology and solid electrolyte membrane, ICBES (International Conference on Biomedical Engineering and Systems) Prague, Czech Republic, August 2014. Ayda Niazi, Carl Anthony, Real time measurement of oxygen by integrating a Clark sensor with printed circuit board technology and solid electrolyte, MNE (International Conference on Micro and Nano Engineering), September 2014. Ayda Niazi, Carl Anthony, Modelling of the oxygen consumption of cells in the cell culturing platform, COMSOL Conference, Cambridge, UK, September 2014. Also a journal paper is submitted and under review: Ayda Niazi, Carl Anthony, Comparison of six miniaturized oxygen sensors, fabricated by printed circuit board technology and solid polymer electrolyte in liquid and gas state measurements. VI Contents Abstract ..................................................................................................................................... II Acknowledgment ...................................................................................................................... V List of Publications .................................................................................................................. VI Contents ................................................................................................................................. VII List of Figures ........................................................................................................................... X List of Tables ........................................................................................................................ XIII 1. Introduction ........................................................................................................................ 1 1.1. Introduction ................................................................................................................. 1 1.2. Aims and Objectives ................................................................................................... 3 1.3. Thesis Overview .......................................................................................................... 5 2. Literature Review ............................................................................................................... 6 2.1. Introduction ................................................................................................................. 6 2.2. Biosensors ................................................................................................................... 6 2.3. Classification of Oxygen Biosensors .......................................................................... 8 2.3.1. Optical oxygen sensors ........................................................................................ 8 2.3.2. Electrochemical oxygen sensors ........................................................................ 10 2.3.2.1. Clark-type oxygen sensor theory of operation ........................................... 12 2.4. Electrochemistry of Clark type oxygen sensor ......................................................... 15 2.4.1. Electrodes ........................................................................................................... 18 2.4.1.1. Working Electrode ...................................................................................... 18 2.4.1.2. Reference Electrode .................................................................................... 18 2.4.1.3. Counter electrode ........................................................................................ 19 2.4.2. Gas Permeable Membrane (Teflon membrane) ................................................. 19 2.4.3. Electrolyte .......................................................................................................... 20 2.4.3.1. Solid Polymer Electrolyte ........................................................................... 20 2.5. Electrical Double Layer ............................................................................................ 22 2.6. Transport Process ...................................................................................................... 24 2.6.1. Diffusion of planar (rectangular) electrodes ...................................................... 27 2.6.2. Diffusion of spherical electrodes ....................................................................... 28 2.6.3. Diffusion of microdisc electrodes ...................................................................... 29 2.7. Kinetics of electrode reactions .................................................................................. 31 2.7.1. Heterogeneous electrochemical reactions .......................................................... 32 2.8. Electrochemical measurement techniques ...............................................................
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