Facilitated transport of Rhodamine B through a novel polymer inclusion membrane (PIM) containing CdTeSe Quantum Dots (QDs) as a carrier reagent has been investigated. The prepared PIM was characterized with Fourier transform infrared (FT-IR) spectroscopy, Atomic Force Microscopy (AFM), confocal microscopy, X-ray Photoelectron Spectroscopy (XPS) and fluorescence spectroscopy (PL) techniques and contact angle measurements. The transport was observed by the increased absorbance peak of the stripping phase by UV-visible and fluorescence spectrophotometer. The transport efficiency of Rhodamine B was observed to be 60% after 6 h under optimized conditions. When the feed phase contained 0.005% Rhodamine B at pH 12.19, Rhodamine B was transported through the PIM (prepared with 0.1% CdTeSe QDs) by using 1 M HCl as a stripping phase. The results also showed that the transport efficiency of the PIM was reproducible and it could be beneficial in long-term separation processes.