Flow Cytometric Measurement Of ROS Production In Macrophages In Response To FcγR Cross-linking

Measurement of reactive oxygen species or ROS is notoriously problematic. In this video, we will demonstrate the reproducible measurement of ROS in response to FC-gamma receptor crosslinking using fluorescent probes and flow cytometry. The main advantages of this technique are the reproducibility of the assay and the use of this method with specific antigenic stimuli, not only mitogens or known ROS inducers.

Dysregulated ROS production is central to the development of many diseases such as chronic granulomatous disease or CGD. Accurately measuring ROS could constitute one part of the molecular diagnosis of CGD. Studying FC-mediated ROS production is important in studying immunodeficiencies, such as CGD, but also in studying auto-immune diseases, or neurodegenerative diseases, where too much ROS leads to oxidative stress and inflammation.