Fluorescence in Microscopy: Probes and Applications

This lecture introduces the concept of fluorescence in microscopy, focusing on different types of fluorescence probes such as organic fluorophores (synthetic dyes, fluorescent proteins) and inorganic fluorophores (lanthanides, quantum dots). The instructor explains the characteristics of synthetic dyes used as fluorescent markers, their molecular weight, penetration through cell membranes, and minimal disturbance to attached molecules. Commonly used dyes in biomicroscopy are discussed, including Rhodamine, Fluorescein, Cyanines, and Coumarins. The lecture also covers the characteristics of various fluorophores used in fluorescence microscopy and flow cytometry, such as their excitation and emission wavelengths. Near-infrared dyes with high extinction coefficients and small Stokes shifts are presented, along with their applications in live tissue imaging.