This study aims to investigate the temperature effect on particle size of copper oxide nanofluid produced under optimal parameters of the Arc Spray Nanoparticle Synthesis System (ASNSS) developed in this research. The purpose is to understand the aggregation feature of copper oxide nanofluid in a higher-than-room-temperature environment and to analyze its size change and the motion behavior of suspended nanoparticles. This study employs an ambient temperature controller to maintain the environment temperature within the scope of normal fluid work temperature to obtain data on the change in suspended particles of copper oxide nanofluid under varying temperatures and through change of time. Experimental result shows that the particle size distribution of copper oxide nanofluid changes when the temperature rises due to the slight absorption and aggregation phenomena between particles, and that the change in environmental temperature can accelerate the aggregation of copper oxide nanofluid, which can affect its stability in application. However, the change in particle size distribution will gradually stabilize for a longer duration of constant temperature.