Reactive Oxygen Species Methods and Protocols

This volume provides a varied selection of current methods and molecular probes used to generate, image and quantify ROS production and lipid peroxidation. Chapters guide readers through methods and protocols on in silico modelling of ROS dynamics in biological systems and a breaking proposal. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

1. Introduction to in silico Modeling to Study ROS Dynamics

Jana Schleicher

2. Assessment of ROS Production in Mitochondria of Live Cells

3. Measurement of Superoxide Production in Acute Hypoxia by Fixed-Cell Microscopy

Pablo Hernansanz-Agustíz, Carmen Choya-Foces, and Antonio Martínez-Ruiz

4. A Photodynamic Tool to Promote a Sustained, ROS-Dependent Growth of Human Hair Follicles in ex vivo Culture

María I. Calvo-Sánchez, Sandra Fernández-Martos, and Jesús Espada

5. Singlet Oxygen and Protochlorophyllide Detection in Arabidopsis thaliana

Liangsheng Wang and Tatjana Kleine

6. Characterization of oxidative lipidomics and autophagy induction in Chlamydomonas reinhardtii under abiotic stress

Cristina Ortega-Villasante, Ángel Barón-Sola, Margarita Toledo-Basantes, Flor Martínez, and Luis E. Hernández

7. Fluorescence Detection of Increased Reactive Oxygen Species Levels in Saccharomyces cerevisiae at the Diauxic Shift

Abhishek Sinha and Elah Pick

8. Flow Cytometry and Confocal Microscopy for ROS Evaluation in Fish and Human Spermatozoa

9. Measurement of Reactive Oxygen Species In Semen Samples Using Chemiluminescence

Tânia R. Dias

10. Methods to Unravel Pathways of Reactive Oxygen Species in the Photodynamic Inactivation of Bacteria

Natalia S. Gsponer and Edgardo N. Durantini

11. Revealing ROS Production by Antibiotics and Photosensitizers in Biofilms. A Fluorescence Microscopy Approach

Sol R. Martínez  and Andrés M. Durantini

12. EPR Spectroscopy as a Method for ROS Quantification in Skin

13. Detection of Superoxide Radical in Adherent Living Cells by Electron Paramagnetic Resonance (EPR) Spectroscopy Using Cyclic Nitrones

Kahina Abbas, Nikola Babic, and Fabienne Peyrot

14. Singlet Oxygen Quantum Yield Determination Using Chemical Acceptors

Roger Bresolí-Obach, Joaquim Torra, Renzo P. Zanocco, Antonio L. Zanocco, and Santi Nonell

15. A General Method to Establish the Relative Efficiency of Different Sonosensitizers to Generate ROS for SDT

Viviana Grosso, Alejandro Senz, and Arnaldo T. Soltermann

16. Lipid Peroxidation Assay using BODIPY-phenylbutadiene Probes:

A Methodological Overview

Juan C. Stockert

17. Use of Fluorescent Probes to Detect ROS in Photodynamic Therapy