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Portable Multiplexed Optical Detection for Point-Of-Care

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Portable Multiplexed Optical Detection for Point-Of-Care PORTABLE MULTIPLEXED OPTICAL DETECTION FOR POINT-OF-CARE A dissertation submitted to the Division of Research and Advanced Studies of the University of Cincinnati in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in the School of Electronic and Computing Systems of the College of Engineering and Applied Science 2012 by Li Shen M.S., Southeast University, Nanjing, China, 2008 B.E., Southeast University, Nanjing, China, 2005 Committee Chair: Ian Papautsky, Ph.D. ABSTRACT In this dissertation, a low-cost, portable and user-friendly optical detection system was developed for microfluidic based lab-on-a-chip (LOC) devices. The conventional analytical methods of measuring these devices require expensive benchtop instruments that are not suitable for point-of-care (POC) applications. The optical detection system developed in this work consists of three major components: a broadband emission white LED to excite multiple fluorephores, a wavelength-independent cross-polarization signal isolation scheme, and a CMOS image sensor for signal detection. The combination of the setup enables simultaneous detection of multiple fluorescent samples. As a proof of concept, the system was applied for optical oxygen detection. The oxygen concentration was indicated by the red luminescence emission intensity of platinum octaethylporphyrin (PtOEP). The sensitivity of the oxygen sensor reached ~41, comparable to the ~50 values reported by others using an external spectrometer. To achieve multi-color analysis, CIE 1931 color space based signal conversion technique was developed, being intuitive and user-friendly. This technique was used for analyzing images of pH and urine glucose colorimetric test strips taken by a camera phone. The linear response ranges are 1-12 and 0-60mM for pH and urine glucose, respectively. Finally, the optical detection system was applied for fluorescent micro-particle detection, achieving ~3µm special resolution. In the particle counting test, 98% and 85% accuracy were achieved in static and dynamic conditions, respectively. With further development and optimization, the optical detection system can be integrated into microfluidic LOC systems for POC applications. ACKNOWLEDGMENTS First and foremost, I would like to thank my advisor Dr. Ian Papautsky for his guidance and constant encouragement during this work. He is always patient and ready to provide innovative ideas and suggestions when I get lost or stuck in my project. Besides technical help, Dr. Papautsky has supported me by providing research assistantship as well as motivating and inspiring me all the time during my stay at University of Cincinnati. I feel gratified to be his student. My gratitude is also expressed to my committee professors, Dr. Fred Beyette, Dr. Jason Heikenfeld, Dr. William Heineman, and Dr. David Klotzkin for their insight and suggestions. I would like to thank Dr. Josh Hagen at Air Force Research Laboratory for supporting and providing valuable advice. I would also like to express my appreciation to Jeff Simkins and Ron Flenniken for their help with cleanroom processing. I would like to thank the Air Force Office of Scientific Research, the Human Effectiveness Directorate, the National Science Foundation, and the University of Cincinnati Institute for Nanoscale Science and Technology for funding my research during these years. Many thanks are in order to my fellow students at the Bio Micro Systems Laboratory group: Ali, Preetha, Choi, Mike, Taher, Yun, Jian, Xing, Xiao, Wenjing, Yuguang, Ananda, Nivedita, for their continuous support and friendship over the years. Last but not the least; I would like to thank my parents Yang Shen and Huiqin Zhou, and my wife Wen Li, for being always together with me, supporting me, and loving me. TABLE OF CONTENTS LIST OF FIGURES ..................................................................................................................... viii LIST OF TABLES ......................................................................................................................... xi CHAPTER 1 INTRODUCTION .....................................................................................................1 Motivation ............................................................................................................................3 Scope of work ......................................................................................................................3 Innovation and significance .................................................................................................5 Chapter Summaries ..............................................................................................................5 CHAPTER 2 FABRICATION OF A PORTABLE FLUORESCENCE DETECTION SYSTEM ..........................................................................................................................................7 Signal isolation for lab-on-a-chip devices ...........................................................................8 Signal detection for lab-on-a-chip devices ........................................................................14 System assembly and testing .............................................................................................21 Summary ............................................................................................................................25 CHAPTER 3 SPECTRAL AND SPATIAL RESOLUTION AND APPLICATION TO GAS SENSING ..............................................................................................................................26 Luminescent oxygen sensing .............................................................................................27 Optimization of the oxygen sensor using ratiometric measurement ..................................33 Summary ............................................................................................................................36 CHAPTER 4 ON-SITE COLORIMETRIC DETECTION USING CAMERA PHONE ..............37 Introduction ........................................................................................................................38 Cell Phone as Photodetector ..............................................................................................40 Measurements of pH and glucose ......................................................................................46 Ambient Light Compensation ............................................................................................49 Summary ............................................................................................................................52 vi CHAPTER 5 STATIC AND DYNAMIC DETECTION OF PARTICLES IN MICROCHANNELS .....................................................................................................................55 Introduction ........................................................................................................................55 Experimental setup and methods .......................................................................................56 Particle imaging and counting ...........................................................................................58 Summary ............................................................................................................................67 CHAPTER 6 CONCLUSIONS .....................................................................................................69 Summary ............................................................................................................................69 REFERENCES ..............................................................................................................................72 vii LIST OF FIGURES Figure Page 1. Schematic of the cross-polarization filtering scheme ........................................................11 2. Transmission of two polarizers placed in parallel (0°), 75°, and crossed (90°) orientations. Insets illustrate isolation of optical signal (Rhodamine B emission centered at 625 nm) from high background signal ............................................................12 3. Overall transmittances of the sandwich structures when one of the five materials was clamped between two orthogonally oriented polarizers. The inset describes the light path during the test...............................................................................................13 4. Schematic of a CMOS image sensor in which photodetector array is covered by Bayer filters ........................................................................................................................16 5. Response of the CMOS sensor to 620nm light as compared to a current measured by a silicon photodiode. The gamma correction function of the CMOS sensor distorts the linear response .................................................................................................19 6. Effect of AWB to the image color and the RGB intensities ..............................................20 7. Responsivity of the CMOS detector in each of its color channels. Inset images taken by the CMOS detector illustrate response at center wavelengths 460 nm, 540 nm and 620 nm............................................................................................................21 8. Schematic of the CMOS image sensor based fluorescence detection system. A broadband white LED was used as excitation and two orthogonally oriented
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