A basket-type fluorescent calix[4]azacrown ether derivative (C[4]azacrown) was conceived and prepared to starting from NBD-chloride (4-chloro-7-nitro-benzofurazan), as a selective turn-off sensor of multi-analytes. The sensor designed as both an useful basket macrocycle and a fluorescent material was characterized by several techniques such as fluorescence spectroscopy, elemental analysis, NMR, FT-IR, and. The optical properties of sensor C[4]azacrown developed for multi-analytes were examined in the presence of anions and cations such as Na+, Ca2+, Mg2+, Fe3+, Fe2+, Ni2+, Co2+, Zn2+, Cu2+, Hg2+, Cr3+, Ag+, Al3+, and F−, Br−, Cl−, H2PO4−, I−, PO43−, SO42−, HSO 4−, CH3COO−, NO3− and several ion/sensor ratios. Nano-basket fluorescent molecule, C[4]azacrown, has a fluorescence response with “on–off” mechanism towards CH3COO−, F− anions and Cu2+ cation with clear selectivity in aqueous medium (H2O/CH3CN 1/4, v/v). The fluorescence properties of C[4]azacrown was also demonstrated by using a confocal microscope in the human lung cancer cell line (A549). It was observed that the fluorescence intensity of the basket sensor quenched clearly in cells performed with Cu2+ ion, CH3COO−, and F− ions, respectively. Moreover, the detection of CH3COO−, F− and Cu2+ ions were performed in grape juice as a food sample. As a result, the basket-type macrocycle, C[4]azacrown, may be performed to recognize Cu2+, CH3COO− and F− ions in living cells and real samples.