Fluorescence in Microscopy: Important Concepts

This lecture covers the fundamental concepts of fluorescence in microscopy, including important terms like excitation and emission spectra, Stokes shift, extinction coefficient, quantum efficiency, brightness, fluorescence lifetime, blinking, quenching, and photobleaching. It also discusses characteristics of fluorescent dyes, fluorophores, and the cycle of a fluorophore. The instructor explains the photobleaching process and its impact on different fluorescent dyes, emphasizing the importance of photostability. Various properties of novel fluorescent protein variants are presented, highlighting factors such as excitation and emission maxima, extinction coefficient, quantum yield, brightness, and photobleaching resistance.