The CBRN (chemical, biological, radiological and nuclear) materials caused to serious health risks that the effective detection of these agents is very important. Macrocyclic fluorescent sensors based on Pillar[5]arene bearing multi Bodipy units have been designed for toxic agents in the CBRN area. The prepared non-symmetric Pillar[5]arene derivative (Pillar[5]-Bodipys) was characterized by NMR, elemental analysis, FT-IR and melting point methods. The macrocyclic sensor exhibits a strong emission around 510 nm, which is used to monitor the interaction of toxic anions. Ion-binding experiments of the chemosensor have been examined in half-aqueous medium with a wide range of anions and the detection process is monitored by UV–vis and fluorescence spectroscopies. The results demonstrated that the chemosensor, Pillar[5]-Bodipys, has a sensitivity and selectivity toward toxic F- ion. The fluorescence intensity of Pillar[5]-Bodipys quenched significantly in the presence of F- anion and LOD was determined as 2.1x10-7 M. In the detection of F- ion, Pillar[5]-Bodipys has also an ideal response time and good stability in a range of pH's. The biological imaging was examined for the detection of F- ion by Pillar[5]-Bodipys, that the fluorescent macrocyclic sensor has a low toxicity. It can be successfully used in the recognition of F- ions on living cells.