How do anti-fading agents work?

Anti-fading agents are substances used in fluorescence microscopy to reduce or prevent photobleaching of fluorescent dyes. Photobleaching occurs when fluorescent dyes or proteins lose their fluorescence. There are several different types of anti-fading agents, each with their own mechanism of action. Most antifade reagents are reactive oxygen species scavengers. Oxygen reacts with fluorescent dyes and proteins, causing photobleaching. Reactive oxygen species scavengers minimize this effect by reacting with and eliminating oxygen from the sample. Some antifade reagents are free radical scavengers. They neutralize free radicals that react with fluorescent dyes and proteins causing photobleaching. Antioxidants work to reduce photobleaching by neutralizing reactive oxygen species (ROS), which prevents the formation of free radicals that cause fading. By reducing photobleaching, these agents help researchers obtain more accurate and reliable imaging of biological samples over time.