The high amounts of oxidants and heavy metal ions caused undesirable health results and the detection of these ions gained increasing importance with new techniques. This paper focuses on a fluorescent macrocyclic sensor based on Pillar[5]arene and Bodipy (Bodipy-P[5]arene) for the recognition of oxidants and heavy metal ions in aqueous solution. After the characterization of the macrocyclic ligand, the spectroscopic measurements were carried out by Uv–vis and fluorescence spectroscopies. The bifunctional fluorescent macrocycle, Bodipy-P[5]arene, presents to a dual effect with fluorescence quenching response for fluorometrically recognizing MnO4- and Hg2+ ions. Bodipy-P[5]arene has low detection limits of MnO4- and Hg2+ such as 1.15 × 10-7 M and 8.12 × 10-7 M, respectively. Moreover, both Bodipy-P[5]arene can respectively detect the target ions in acidic and basic mediums and the macrocyclic ligand has short response times. In addition, the Bodipy-P[5]arene probe demonstrates low cellular toxicity and exceptional membrane permeability, making it a successful tool for fluorescently imaging Hg2+ and MnO4- ions within living HeLa cells.