Iowa State University Capstones, Theses and Graduate Theses and Dissertations Dissertations 2014 Development of matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI- MSI) for plant metabolite analysis Andrew Korte Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/etd Part of the Analytical Chemistry Commons Recommended Citation Korte, Andrew, "Development of matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) for plant metabolite analysis" (2014). Graduate Theses and Dissertations. 14021. https://lib.dr.iastate.edu/etd/14021 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Graduate Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Development of matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) for plant metabolite analysis by Andrew R. Korte A dissertation submitted to the graduate faculty in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Major: Analytical Chemistry Program of Study Committee: Young-Jin Lee, Major Professor R. Sam Houk Basil Nikolau Emily Smith Patricia Thiel Iowa State University Ames, Iowa 2014 . ii TABLE OF CONTENTS ACKNOWLEDGMENTS ......................................................................................................iv ABSTRACT ........................................................................................................................ v CHAPTER 1: GENERAL INTRODUCTION .................................................................................... 1 Background ............................................................................................................................ 1 MALDI-MSI: General Workflow ............................................................................................. 4 Dissertation Organization ...................................................................................................... 5 Figure ..................................................................................................................................... 7 CHAPTER 2: MASS SPECTROMETRIC IMAGING AS A HIGH-SPATIAL RESOLUTION TOOL FOR FUNCTIONAL GENOMICS: TISSUE-SPECIFIC EXPRESSION OF TT7 INFERRED FROM HETEROGENEOUS DISTRIBUTION OF METABOLITES IN ARABIDOPSIS FLOWERS ................................................................................................................................... 8 Abstract ................................................................................................................................. 8 Introduction ........................................................................................................................... 9 Experimental ....................................................................................................................... 12 Results and Discussion ........................................................................................................ 16 Conclusions .......................................................................................................................... 23 Acknowledgment................................................................................................................. 24 Figures ................................................................................................................................. 25 Supplemental Figures .......................................................................................................... 31 CHAPTER 3: MALDI-MS ANALYSIS AND IMAGING OF SMALL MOLECULE METABOLITES WITH 1,5-DIAMINONAPHTHALENE (DAN) ............................................................................ 36 Abstract ............................................................................................................................... 36 Introduction ......................................................................................................................... 36 Experimental ....................................................................................................................... 38 Results and Discussion ........................................................................................................ 42 Conclusions .......................................................................................................................... 46 Acknowledgment................................................................................................................. 47 Figures ................................................................................................................................. 48 Supplemental Tables and Figures ....................................................................................... 52 CHAPTER 4: MULTIPLEX MASS SPECTROMETRIC IMAGING WITH POLARITY SWITCHING FOR CONCURRENT ACQUISITION OF POSITIVE AND NEGATIVE ION IMAGES ..................... 57 Abstract ............................................................................................................................... 57 Introduction ......................................................................................................................... 58 Experimental ....................................................................................................................... 60 Results and Discussion ........................................................................................................ 62 Conclusions .......................................................................................................................... 67 Acknowledgments ............................................................................................................... 69 iii Figures ................................................................................................................................. 70 Supplemental Figure ........................................................................................................... 74 CHAPTER 5: SINGLE-CELL MALDI-MS IMAGING OF CORN LEAF METABOLITES BY MODIFICATION OF MALDI-LTQ-ORBITRAP DISCOVERY OPTICS ........................................... 75 Abstract ............................................................................................................................... 75 Introduction ......................................................................................................................... 75 Experimental ....................................................................................................................... 77 Results and Discussion ........................................................................................................ 81 Conclusions .......................................................................................................................... 85 Acknowledgment................................................................................................................. 86 Figures ................................................................................................................................. 87 Supplemental Table and Figures ......................................................................................... 92 CHAPTER 6: SUMMARY AND OUTLOOK ................................................................................ 94 Summary ............................................................................................................................. 94 Outlook ................................................................................................................................ 94 APPENDIX: MULTIPLEX MALDI-MS IMAGING OF PLANT METABOLITES USING A HYBRID MS SYSTEM ................................................................................................ 97 Abstract ............................................................................................................................... 97 Introduction ......................................................................................................................... 98 Materials .............................................................................................................................. 99 Methods ............................................................................................................................ 102 Notes ................................................................................................................................. 110 Acknowledgments ............................................................................................................. 115 Figures ............................................................................................................................... 116 REFERENCES .......................................................................................................................... 120 iv ACKNOWLEDGMENTS I am greatly indebted to Dr. Young-Jin Lee, who in addition to offering guidance and mentorship as my adviser provided me the opportunity to work on challenging and exciting projects. I would also like to acknowledge my committee members, Dr. Sam Houk, Dr. Basil Nikolau, Dr. Emily Smith, and Dr. Patricia Thiel for their guidance