Two novel bis naphthaldehyde functionalized fluorophores, HF-NAP1 and HF-NAP2, were designed and synthesized using a 4,4-(hexafluoroisopropylidene)dianiline core. Structural alteration at the ortho position adjacent to the imine moiety enabled control over Excited-state intramolecular proton transfer (ESIPT) phenomenon. HFNAP1 bearing an ortho-hydroxyl group exhibited ESIPT activity, while HF-NAP2 with an ortho-methoxy substituent suppressed this process. Both molecules exhibited two distinct emission colors (HF-NAP1 emitted green and HF-NAP2 emitted blue fluorescence) with aggregation-induced emission (AIE) characteristics in THF-water mixtures. Single-crystal X-ray diffraction analysis confirmed molecular geometry, while solvent dependent absorption and emission studies established ESIPT contributions. Fluorescence lifetime, DLS, SEM, TEM, and quantum yield analyses further validated aggregation driven restriction of intramolecular motion. The detailed density functional theory (DFT) and time-dependent DFT calculations supported the keto-enol tautomerism and electronic transitions of the molecules. Finally, bioimaging experiments in live HeLa cells demonstrated efficient cellular uptake and bright intracellular fluorescence, confirming the potential of HF-NAP1 and HF-NAP2 as next generation ESIPT/AIE active probes for biomedical imaging.