In the present work, we report the design of a novel chemosensor 2-(1H-indole-3-yl)- 1H-imidazo[4,5-b]phenazine, (R) for fluoride ion detection via a naked eye observable colour change from light yellow to brown along with 10 fold ratiometric fluorescence quenching from green to no fluorescence in a matrix containing a large number of similar toxic anions in aqueous dimethylsulfoxide (30% H 2 O + DMSO) solution. The sensing mechanism was ascertained to be the fluoride triggered deprotonation of imidazole as well as indole moiety of R which is supported by 1:2 saturation level equivalence of R-F -. R was fully characterized through FT-IR, 1 H, 13 C NMR and HRMS. R is an effective receptor for Fwithin a wide range of pH that indicates its bio-compatibility. LoD and K a for Fwere found to be 1.8 × 10 -6 M (below 0.34 mg/Liter) and 2.7 × 10 4 M -1 respectively. The experimental observations were supported by the theoretical calculations using density functional theory. R also showed its potential practical applicability for fluoride detection in toothpaste solution. Furthermore, to prove its nontoxicity, probe R also applied to a cytotoxicity assay of HeLa cells followed by confocal fluorescence cells imaging and Zebrafish imaging experiments.[a] S. Naha, Prof. S. Velmathi