Introduction: Due to very small dimensions, nano-materials can have different properties compared to their bulk form. The nanoparticle size dominates some of the atom-related properties of nanoparticles. Such behavior changes may adversely affect biological organism, especially phytoplankton as the basis of food chain cycle. For this reason, in current research, the toxicity effect of CuO nanoparticles (CuO NP) on Scenedesmus dimorphus has been evaluated. Materials and Methods: The toxic effect of CuO NP was investigated at 5 different concentrations of 2. 5, 6. 5, 17. 4, 45. 7, 120 mg/L against control in three replicates for each treatment and control. All samples were treated at constant temperature of 24 ° C and 12-hour period of light and dark for 72 hours. Algae count was done at time intervals of 24, 48 and 72h. Chlorophyll assay was done according to ASTM standard. Results: EC10, EC50, and EC90 amounts for 72 hours were 0. 18, 28. 84 and 4477. 35 mg/L, respectively. Over time, chlorophyll concentration was decreased in all treatments except for 57. 68 mg/L. Carotenoid showed significant decrease between all treatments and the control and also among different treatments (p<0. 05) and in 72h three treatments had different carotenoids (p<0. 05). Discussion and Conclusion: These results showed that CuO NP had significant toxic effect on Scenedesmus dimorphus and resulted in reduction, growth rate, and increase in doubling time. Also, the percent inhibition was increased with increasing CuO NP concentration. The chlorophyll and carotenoid levels decreased in Scenedesmus dimorphus by increasing CuO NP concentration.