In the mid 1990s, fluorescence was considered as an old technique, not worth a great deal of resources and effort. Nonetheless, it continued to be used in classical fashion, as well as truly innotative ways. Over the past few years, fluorescence has gained in popularity, as many of the new approaches have become more widely known and appreciated. Given their many advantages, fluorescence approaches are expected to continue to be applied in the fields of biophysics and biophysical chemistry for a very long time.