The effective detection of lanthanide ions is very important due to the penetration of these ions to the living cells. In this paper, novel Pillar[5]arenes, as a new fascinating class supramolecular chemistry, including ten and five nitrobenzofurazan (NBD) fragments were prepared to examine their optical behavior towards trivalent lanthanide ions. The Pillar[5]arene derivatives including ten or five NBD units were named as d-NBD-P5 and p-NBD-P5, respectively. The target macrocyclic compounds were characterized by some techniques such as NMR, melting point, FT-IR, elemental analysis and mass spectroscopy. The response of the synthesized probes towards trivalent lanthanide ions was fluorometrically examined with fluorescence spectroscopy in a half aqueous medium that the probes have especially sensitivity for two metal ions, Eu (III) and Yb (III). The confocal microscope was also used to support visually the sensitivity of d-NBD-P5 and p-NBD-P5 towards target lanthanide ions on living cells. Thus, the results obtained from the solution medium were also supported in the cell medium and both macrocyclic compounds have low toxicity.