A NOVEL OPTICAL APPROACH TO THE INTRAOPERATIVE DETECTION OF PARATHYROID GLANDS By Constantine A Paras Dissertation Submitted to the Faculty of the Graduate School of Vanderbilt University in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biomedical Engineering May, 2012 Nashville, Tennessee Approved: Professor Anita Mahadevan-Jansen (chair) Professor Duco Jansen Professor Robert Galloway Dr. James Broome Dr. Edward M. Brown Copyright 2012 by Anita Mahadevan-Jansen and Constantine A. Paras All Rights Reserved To Dean TABLE OF CONTENTS ................................................................................................................................................. PAGE LIST OF TABLES ......................................................................................................................... vi LIST OF FIGURES ...................................................................................................................... vii CHAPTER 1.Introduction .................................................................................................................................. 1 Motivation ................................................................................................................................... 2 Specific Aims .............................................................................................................................. 4 Specific Aim (1): ..................................................................................................................... 4 Specific Aim (2): ..................................................................................................................... 5 Specific Aim (3): ..................................................................................................................... 5 2. BACKGROUND AND SIGNIFICANCE .................................................................................. 7 Anatomy of the Neck .................................................................................................................. 8 Diseases in the neck .................................................................................................................. 10 Current localization techniques ................................................................................................. 13 Optical Spectroscopy................................................................................................................. 15 Fluorescence Spectroscopy ....................................................................................................... 18 Previous Research ..................................................................................................................... 22 In vitro studies ....................................................................................................................... 22 In vivo Raman study .............................................................................................................. 25 Summary ................................................................................................................................... 27 References ................................................................................................................................. 27 3.NEAR-INFRARED AUTOFLUORESCENCE FOR THE DETECTION OF PARATHYROID GLANDS ...................................................................................................................................... 33 Introduction ............................................................................................................................... 34 Methods ..................................................................................................................................... 35 Results ....................................................................................................................................... 37 Discussion ................................................................................................................................. 39 Conclusion ................................................................................................................................. 42 References ................................................................................................................................. 42 iv 4.UNDERSTANDING THE BASIS OF THE NEAR INFRARED FLUORESCENCE FOUND IN PARATHYROID TISSUES .................................................................................................... 46 Introduction ............................................................................................................................... 47 In vivo NIR fluorescence properties ......................................................................................... 50 Analysis ..................................................................................................................................... 52 Analysis of variance (ANOVA) ............................................................................................ 55 Optical properties ...................................................................................................................... 56 Method ................................................................................................................................... 56 Analysis ................................................................................................................................. 56 EEM ....................................................................................................................................... 57 Molecular basis of NIR fluorescence ........................................................................................ 59 Tissue NIR fluorescence ........................................................................................................ 62 Discussion ................................................................................................................................. 67 Future studies ............................................................................................................................ 71 References ................................................................................................................................. 72 5.DEVELOPMENT OF A REAL-TIME INTRA-OPERATIVE PARATHYROID VISUALIZATION SYSTEM FOR ENDOCRINE SURGERY .................................................. 77 Introduction ............................................................................................................................... 78 Methods and Materials .............................................................................................................. 83 Results ....................................................................................................................................... 89 Conclusions ............................................................................................................................... 93 Recommendations ..................................................................................................................... 94 References ................................................................................................................................. 95 6.CONCLUSIONS........................................................................................................................ 97 Summary of Chapters ................................................................................................................ 98 Future work ............................................................................................................................. 101 BIBLIOGRAPHY ....................................................................................................................... 103 v LIST OF TABLES Chapter 4 Page Table 1: ANOVA Results: Average Parathyroid Intensity ..............................................55 Table 2: Summary of all cases for disease groupings ......................................................63 Table 3: Summary of disease groupings after removing influential observation ............63 Table 3: Ratios of normalized parathyroid signal to thyroid signal and resulting p-values .......................................................................................................................... 64 Table 4: Optical Properties of Tissues in the Neck ..........................................................69 vi LIST OF FIGURES Chapter 2 Figure 1: Anatomy of the central neck (2). Figure 2: Thyroid (blue arrows), and parathyroid (yellow arrows) glands under H&E (1). Figure 3: Jablonski diagram illustrating the various processes including fluorescence. Figure 4: Typical absorption and emission spectra. Figure 5: Fluorescence spectra of parathyroid and thyroid tissues at 337 nm excitation. Figure 6: (a) Image of the clinical Raman system, (b) Schematic of the Raman system. Figure 7: Raman spectra of a typical patient. Chapter 3 Figure 1: (a) Fluorescence spectra measured from parathyroid, thyroid, fat, muscle and trachea of a typical patient. (b) Fluorescence spectra from parathyroid and thyroid tissue normalized to their respective peaks. Figure 2: (a) Average fluorescence peak intensity and (b) normalized peak intensity from parathyroid and thyroid measurements from each patient. Chapter 4 Figure 1: Proposed structural model of the predicted