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Table of contents
ABBREVIATIONS
RÉSUMÉ DE LA THÈSE (FRENCH ABSTRACT)
GENERAL INTRODUCTION
PART A: STATE-OF-THE-ART
1. OXIDATIVE STRESS AND REACTIVE OXYGEN/NITROGEN SPECIES (ROS/RNS)
1.1. Natural process of oxidative stress
1.2. Roles of ROS/RNS in life
1.2.1. Beneficial effects
1.2.2. Deleterious effects
1.3. Conclusion of section 1
2. DETECTION METHODS FOR ROS/RNS
2.1. Some general analytical methods
2.1.1. Fluorescence method
2.1.2. Chemiluminescence method
2.1.3. Electron spin resonance (ESR) spectroscopy
2.1.4. Ultraviolet-visible (UV-Vis) spectroscopy
2.1.5. Biomarkers method
2.2. Electrochemical method
2.2.1. Superoxide anion (O●– 2) sensors
2.2.2. Hydrogen peroxide (H2O2) sensors
2.2.3. Nitric oxide (NO•) sensors
2.2.4. Peroxynitrite (ONOO– ) sensors
2.2.5. Nitrite ion (NO– 2) sensors
2.2.6. Platinum black (Pt-black) coated electrodes
2.3. Conclusion of section 2
3. ANALYTICAL PROGRESS IN EX VIVO DETECTION OF CELLULAR ELECTROACTIVE MESSENGERS
3.1. Single-cell analysis by using microelectrodes
3.1.1. Microelectrodes and their advantages
3.1.2. Detection of electroactive messengers from a single living cell
3.2. Microfluidic devices as functional tools for cell-based analysis
3.2.1. Cell culture in microfluidic devices
3.2.2. Cell manipulation in microfluidic devices
3.2.3. Cell-based analysis on microfluidic platforms
3.3. Conclusion of section 3
4. CONCLUSION OF STATE-OF-THE-ART AND PERSPECTIVES
PART B: IN VITRO ELECTROCHEMICAL DETECTION OF ROS/RNS AT HIGHLY SENSITIVE PT/PT-BLACK MICROBAND ELECTRODE INSIDE MICROFLUIDIC SYSTEM
1. CONVECTIVE-DIFFUSIVE MASS TRANSPORT ABOVE PT/PT-BLACK MICROCHANNEL ELECTRODE
1.1. Mass transport regimes above a single microband electrode
1.2. Choice of mass transport regime in our microfluidic device
1.3. Optimization and evaluation of Pt-black deposit
1.4. Conclusion of section 1
2. ROS/RNS IN VITRO DETECTION AT PT/PT-BLACK MICROBAND ELECTRODE
2.1. In vitro detection of two stable candidates: H2O2 and NO– 2
2.1.1. Current responses at Pt/Pt-black electrode
2.1.2. Oxidation mechanisms of H2O2 and NO– 2
2.1.3. Calibration curve and electrode sensitivity
2.2. In vitro detection of two unstable candidates: ONOO– and NO•
2.2.1. Detection of ONOO–
2.2.2. Detection of NO•
2.3. In vitro detection of samples mixture
2.4. Conclusion of section 2
3. CONCLUSION OF PART B
PART C: MONITORING OF CELLULAR OXIDATIVE STRESS BY PT/PT-BLACK ELECTRODES-INTEGRATED MICROSYSTEMS
1. INVESTIGATION OF OXIDATIVE STRESS IN LABORATORY
1.1. Cell models
1.2. Cell stimulations
1.2.1. Mechanical penetration
1.2.2. Biochemical stimulation
1.3. Conclusion of section 1
2. DETECTION OF OXIDATIVE BURSTS FROM CELLS POPULATIONS ON BASAL MICROBAND ELECTRODE
2.1. Experimental conditions
2.1.1. Microdevice configuration
2.1.2. Cells viability and density
2.2. Qualitative detection of ROS and RNS production
2.2.1. Characteristics of amperometric responses
2.2.2. Cells variations and successive releases
2.2.3. Cells detachment and chip reuse
2.3. Quantification of ROS and RNS production
2.3.1. Evaluation of fluxes and quantities of ROS and RNS production
2.3.2. Evaluation of primary production of O●– 2 and NO•
2.4. Conclusion of section 2
3. DETECTION OF OXIDATIVE BURSTS FROM CELLS POPULATION ON DOWNSTREAM MICROBAND ELECTRODES
3.1. Experimental conditions
3.1.1. Microdevice configuration
3.1.2. Cells manipulation inside microdevice
3.2. Detection of ROS and RNS production
3.2.1. Detection reproducibility between parallel electrodes
3.2.2. Detection reliability after biocompatible coating
3.2.3. Detection during continuous flow
3.2.4. Detection after 10-min stop flow
3.3. Conclusion of section 3
4. CONCLUSION OF PART C
GENERAL CONCLUSION AND PERSPECTIVES
APPENDIX
APPENDIX I. ELECTROCHEMICAL PRINCIPLES AND MICROELECTRODES
I.1. Electrochemical principles
I.2. General electrochemical techniques
I.3. Microelectrodes and their electrochemical performances
APPENDIX II. MICROFLUIDIC MATERIALS AND MICROFABRICATION
II.1. Materials
II.2. Microfabrication techniques
APPENDIX III. EXPERIMENTAL SECTION
III.1. Solutions preparation
III.2. Sensors preparation
III.3. Raw 264.7 macrophages preparation
III.4. Analytical measurements
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