Fluorescein molecules are extensively used to develop fluorescent probes for various analytes due to their excellent photophysical properties and the spirocyclic structure. The main structural modification of fluorescein occurs at the carboxyl group where different groups can be easily introduced to produce the spirolactam structure which is non-fluorescent. The spirolactam ring opening accounts for the fluorescence and the dual sensing of analytes using fluorescent sensors is still a topic of high interest. There is an increase in the number of dual sensors developed in the past five years and quite a good number of fluorescein derivatives were also reported based on reversible mechanisms. This review analyses environmentally and biologically important cations such as Cu2+, Hg2+, Fe3+, Pd2+, Zn2+, Cd2+, and Mg2+; anions (F−, OCl−) and small molecules (thiols, CO and H2S). Structural modifications, binding mechanisms, different strategies and a comparative study for selected cations, anions and molecules are outlined in the article.