This study aims to evaluate the effect of ionic liquid intercalation in smectite on fluorescent dye adsorption. In the first step, we investigated the influence of interlayer cation of Montmorillonite (Li+, Na+, K+, Rb+, and Cs+) on the intercalation of 1-butyl-3-methylimidazolium chloride. The adsorption data was well described by the pseudo-second-order kinetics and Langmuir adsorption isotherm. The material structure was investigated by Fourier transform infrared (FTIR) and wide-angle X-ray diffraction (XRD) showing that the ionic liquid intercalation is occurred. Then, the organo-clays were used to the adsorption of a fluorescein. In presence of ionic liquid the dye adsorption is a little bit improved. The interaction between the dye and the ionic liquid are highlighted based on the fluorescence lifetime. This interesting result confirms that the dye is not released compared to the pristine Montmorillonite.