A multi-layer of the dye, natural colorant Berberine, was successfully developed by the self-assembly deposition from water-soluble cationic dye(Berberine chloride) and anionic polyelectrolyte PSS(Polysodium 4-styrenesulfonate) in aqueous solution via electrostatic attraction. The corresponding results on multi-layer were characterized by UV-Vis absorbance measurements. The growth of multi-layer formed by the sequential interaction was also determined. The findings measured by UV-Vis spectrophotometer showed that the bilayer deposition characteristic was linear and highly reproducible from layer to layer.