The construction of multifunctional supramolecular liquid crystals is a hot topic in current research. Herein, we reported the strategy of acylhydrazone-functionalization to construct a novel series of triphenylamine-based multifunctional V-shaped liquid crystals (namely, TPA-n). These V-shaped molecules can not only self-assemble into room-temperature hexagonal columnar phase over a very wide range of temperatures, but also form fluorescent gels with the morphology of intertwined fibers in n-heptanol. Furthermore, based on the chelation-enhanced fluorescence quenching (CHEQ) mechanism, TPA-n demonstrated excellent fluorescent probes properties for rapid (respond time < 10 s), highly selective and sensitive (LOD = 0.72 nM) detection of Cu2+ in a mixed aqueous solution. Most importantly, probe TPA-12 has been successfully applied for the quantity analysis of Cu2+ in real water samples and fluorescent imaging of Cu2+ in live HT-22 cells.