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• A phenolphthalein-aminophenol (PFAP)-based chemosensor for Al3+ was synthesized. • PFAP showed a LOD of 1.5 µM and a fast response time of 10 s. • PFAP was used to detect Al3+ in the intracellular region of the A549 cancer cells. Monitoring of excess Al3+ and its selective detection in biological and environmental samples are important tasks for researcher due to its harmful effects. Herein, we present a phenolphthalein-aminophenol derived sensor (PFAP) for the selective fluorescent response of Al3+ in an "OFF-ON" mode over a pool of analytes including eighteen cations with striking greenish emission. It has a prominent limit of detection (LOD) value (1.5 µM), fast response time (10 s) for Al3+ detection. The complexation properties of PFAP with Al3+ ions were clarified by UV–vis, 1H & 13C NMR, HRMS, and FTIR spectroscopic experiments. The recognition mechanism of PFAP for Al3+working with chelation enhanced fluorescence (CHEF) and is verified with two model compounds PF and PFAN in the light of UV–vis, PL, TRPL, and NMR experiment. To check its applicability, easily prepared test paper and TLC strip of PFAP was produced for rapid and selective onsite detection of Al3+ ions. Bio-imaging application of PFAP in human lung cancer cell lines A549 demonstrated excellent results with negligible cytotoxicity and as an excellent marker to detect traces of Al3+ ion in a time-dependent as well as concentration-dependent manner. Actual sample analysis for Al3+ with the probe PFAP produces fruitful result. [Display omitted] [ABSTRACT FROM AUTHOR]