High-content live cell imaging of ROS production in activated macrophages
Immune cells play a role in maintaining a normal body by eliminating foreign substances through the inflammatory response. On the other hand, excessive inflammation can cause autoimmune diseases, allergies, diabetes, and Alzheimer‘s disease. Monocytes stimulated by foreign substances produce reactive oxygen species (ROS) in mitochondria, differentiate into activated macrophages, and decompose foreign substances. ROS have important functions as signaling molecules, but can also be toxic byproducts that damage the DNA within cells. Fluorescent excitation also produces ROS, causing phototoxicity issues that damage living cells. This application note introduces low-phototoxicity, high content live-cell imaging for quantitative analysis of ROS by detecting ROS in activated macrophages using weak excitation light and a highly sensitive monochrome camera and performing label-free counting on cells within the field of view.