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High-Performance Electrochemical Detection of Reactive Oxygen High-performance electrochemical detection of reactive oxygen/nitrogen species inside microfluidic devices : application for monitoring oxidative stress from living cells Yun Li To cite this version: Yun Li. High-performance electrochemical detection of reactive oxygen/nitrogen species inside mi- crofluidic devices : application for monitoring oxidative stress from living cells. Chemical Physics [physics.chem-ph]. Université Pierre et Marie Curie - Paris VI, 2014. English. NNT : 2014PA066367. tel-01133304 HAL Id: tel-01133304 https://tel.archives-ouvertes.fr/tel-01133304 Submitted on 19 Mar 2015 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Université Pierre et Marie Curie Ecole doctorale de Chimie Physique et Chimie Analytique de Paris Centre ED 388 UMR CNRS 8640 PASTEUR, Groupe d’Electrochimie High-Performance Electrochemical Detection of Reactive Oxygen/Nitrogen Species inside Microfluidic Devices. Application for Monitoring Oxidative Stress from Living Cells. Par Yun LI Thèse de doctorat d’Electrochimie Présentée et soutenue publiquement le 17 septembre 2014 devant le jury composé de : Dr. Fethi BEDIOUI (ENSCP) Rapporteur Pr. Pierre GROS (Université de Toulouse) Rapporteur Pr. Didier DEVILLIERS (UPMC) Examinateur Pr. Bruno LE PIOUFLE (ENS Cachan) Examinateur Dr. Laurent THOUIN (ENS) Co-directeur de thèse Dr. Christian AMATORE (ENS) Directeur de thèse Acknowledgements With the support of China Scholarship Council, I have undergone a great experience in pursuing my academic dreams as a PhD candidate in ENS, running like a river, small at first and then passionately past boulders and over waterfalls. During this period, many people behind the scene have made significant contributions, in one way or another, and hereby I sincerely express my heartfelt gratitude. First and foremost, my great appreciation goes to Mr. Christian Amatore, my thesis director, for offering me such a chance to work in his electrochemistry group. Not only his intelligence or morality, but also the passion toward science, benefits me and makes me a fortunate child of favorable circumstance. Special thanks for his confidence and his effort of helping me to find a great postdoc position in Mirkin’s group (CUNY, USA) to continue the study in electrochemistry. My sincere thanks also go to Mr. Fethi Bedioui and Pierre Gros who have kindly accepted to evaluate this work and compose a relevant report to UPMC. The equivalent appreciation is own to Mr. Didier Devilliers and Bruno Lepioufle for acceding to examine this thesis. This thesis is accomplished under supervision of Laurent Thouin. But for him, I could not have made such a progress in accumulation of knowledge. Thanks for his rigorous attitude, sense of responsibility, and degree of sophistication to view issues from different perspectives. Meanwhile, I am truly grateful to Catherine Sella for her unceasing effort to help and extreme patience during our discussion. Many thanks go to Frédéric Lemaître and Manon Guille-Collignon for their constant concern and help in biological realm. Their brilliance as well as diligent guidance has considerably broadened my horizon, correspondingly inspiring me to extend the interest toward bioanalytical domain. I would also like to personally thank my previous supervisors in China: Xinghua Xia and Weihua Huang. It is they who have inspired my academic voyage, leading me to a magnificent life. Thanks to my colleagues in ENS: Cong (share joy and sorrow, be my support all the time); Wided and Pierre (many memories in the lab); Pierluca (for plenty of encouragements); Anne and Rémy (help me to become gradually familiar with microfabrication and manipulation of cells); Thomas, Ana, Xiaoqing, Guillaume, Lylian and Dali (make the scientific life very interesting and active); Feli, Margherita, Jing Zhu, etc. The colorful life will be forever embedded in my memory. Thanks to Yong Chen and the people in his group: Megan, Xin Li, Junjun Li, Jiaji Liu, Zhitao Han, Fan Zhang, Jian Shi, Jun Liu, Jie Hu, Sisi Li, Wentao He, Zhi Li, Yu Zhang and Yadong Tang, for their help in microfluidic fabrication and company in leisure time. Thanks to Marie-Aude and Kalthoum for the generous help in cell culture. Thanks to Fatima, Raquel and Wei Wang for giving me a lot of perceptive advice in this work. Great thanks to Cécile (impressively capable and hardworking), Géradine and Eric for the smooth and orderly running of all laboratory facilities. I shall extend my thanks to Dominique Ho Tin Noe, Anne Halloppe, Frédéric Bataille, Auguste Filippi in ENS; Soobrayen Koonavadee and Hélène Gérard in UPMC. They are always so nice to help me arrange the multifarious student affairs. Finally, I would like to express my deepest gratitude to my family and friends. Thanks Dad, for setting up a great example of perseverance, and for his incessant encouragements in my frustrations. Thanks Mom, for not only being a tender mother, but also my best friend. Thanks to my friends Hao Chen, Cheng Qian, Yinzi Cao, Jing Li, Jun Dong, Yuan Hu, Jiazhen Zhu, Xiangxue Kong, Liang Kong, Chunjuan Sheng, Xing Jin, Lin Lin, Yi Peng, etc. They are always my courage through the exciting but sometimes arduous journey. Table of contents ABBREVIATIONS ................................................................................................. 1 RÉSUMÉ DE LA THÈSE (FRENCH ABSTRACT) ........................................... 7 GENERAL INTRODUCTION ............................................................................ 35 PART A: STATE-OF-THE-ART 1. OXIDATIVE STRESS AND REACTIVE OXYGEN/NITROGEN SPECIES (ROS/RNS) ...................... 41 1.1. Natural process of oxidative stress .............................................................................................. 41 1.2. Roles of ROS/RNS in life .............................................................................................................. 46 1.2.1. Beneficial effects ............................................................................................................. 47 1.2.2. Deleterious effects ........................................................................................................... 49 1.3. Conclusion of section 1 ................................................................................................................ 50 2. DETECTION METHODS FOR ROS/RNS .................................................................................... 51 2.1. Some general analytical methods ................................................................................................ 51 2.1.1. Fluorescence method ....................................................................................................... 51 2.1.2. Chemiluminescence method ........................................................................................... 53 2.1.3. Electron spin resonance (ESR) spectroscopy .................................................................. 55 2.1.4. Ultraviolet-visible (UV-Vis) spectroscopy ..................................................................... 56 2.1.5. Biomarkers method ......................................................................................................... 57 2.2. Electrochemical method............................................................................................................... 58 ●– 2.2.1. Superoxide anion (O 2) sensors ....................................................................................... 60 2.2.2. Hydrogen peroxide (H2O2) sensors ................................................................................. 61 2.2.3. Nitric oxide (NO•) sensors .............................................................................................. 63 – 2.2.4. Peroxynitrite (ONOO ) sensors ....................................................................................... 66 – 2.2.5. Nitrite ion (NO2 ) sensors ................................................................................................ 67 2.2.6. Platinum black (Pt-black) coated electrodes ................................................................... 69 2.3. Conclusion of section 2 ................................................................................................................ 70 3. ANALYTICAL PROGRESS IN EX VIVO DETECTION OF CELLULAR ELECTROACTIVE MESSENGERS............................................................................................................................. 71 3.1. Single-cell analysis by using microelectrodes ............................................................................. 72 3.1.1. Microelectrodes and their advantages ............................................................................. 72 3.1.2. Detection of electroactive messengers from a single living cell ..................................... 73 3.2. Microfluidic devices as functional tools for cell-based analysis ................................................. 77 3.2.1. Cell culture in microfluidic devices ...............................................................................
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