Two novel highly selective fluorescent sensors L‐1 and L‐2 for the determination of Al3+ ion were synthesized and characterized. The sensors acridine‐4,5‐diylbismethyl(bis(oxy))bis(2,1‐phenylene))bis‐(methylene))bis(azanediyl))bis(3‐phenylpropan‐1‐ol) (L‐1) and 2,2′‐acridine‐4,5‐diylbis(methylene(bis(oxy))bis(2,1‐phenylene))bis‐(methylene))bis(azanediyl))bis(propan‐1‐ol) (L‐2) showed “turn on” fluorescence response in the presence of Al3+ ion in DMSO/tris‐HCl (1:1, v:v, pH=7.2) buffer solution. Other cations viz. Li+, Na+, K+, Mg2+, Ca2+, Fe2+, Co2+, Ni2+, Cd2+, Cu2+, Zn2+, Mn2+, Ba2+, Pb2+, Hg2+, Fe3+ and Eu3+ ions showed no appreciable change in fluorescence spectrum. The detection limits of L‐1 and L‐2 for Al3+ ion were at the parts per billion level. In addition, possible utilization of sensors as bio‐imaging fluorescent sensors to detect Al3+ in human cervical HeLa cells was also investigated by confocal fluorescence microscopy.