This study aims to check the fluorescence property of acrylic fabrics dyed with Juglans regia bark extract. Fluorescence measurements have been developed on the aqueous extract and acrylic samples. The extraction process was assisted by a microwave with the following conditions: a concentration of 5 g·L−1, a power of 850, a pH of 5 and an extraction duration of 4 min. Afterwards, the dyeing quality was assessed by measuring the color strength (K/S) and the photoluminescence intensity (PL) of acrylic fibers dyed at 350 W for 3 min, with the extract already prepared. The effect of certain factors (pH, power, concentration and duration of dyeing) on the dyeing process was also investigated. Subsequently, this process was optimized thanks to the surface response method in order to maximize the photoluminescence intensity of dyed acrylic fibers. Best dyeing properties were achieved at 500 W, pH 2 for 4 min. The results showed good washing fastness and acceptable light fastness.