In bulk materials, positional isomers not only help in understanding how slight difference in molecular structure alters the crystal packing and optical properties, but also play a key role in developing new type of materials for functional applications. A detailed study on the photophysical properties of fluorene-HBT positional isomers in solution and in the solid state providing a molecular level understanding of the factors which influence fluorescence behavior is reported. Two molecules Ia and IIa were synthesized by Suzuki coupling reaction and their photophysical properties were compared to positional isomers Ib and IIb. Crystal structure analyses and density functional theory (DFT) computation studies were performed to understand structure-properties relation and the results reveal that changing substitution pattern has a marked influence on their packing modes and luminescence properties. Strong noncovalent interactions in the solid state hamper the excited state intramolecular proton transfer (ESIPT) process which causes fluorescence quenching in the solid state (Ia and IIa = φf, 28-40%; Ib and IIb = φf, 55-67%). Compounds show solvent-responsive and aggregation induced emission (AIE) properties. Bent structures of Ia with double and symmetric substitution of ESIPT motifs exhibit particularly unique condensed phase upon heating, confirmed as a nematic liquid crystalline phase, and this is the first report on the ESIPT and AIE active liquid crystalline materials with a banana-shaped molecule.