Different laser parameters and the interactional production can affect the barcode grade profoundly. In order to mark barcodes on aluminum alloy with a laser effectively, it is important to search out the optimum parameters and establish a relationship between critical laser parameters and the quality of the symbol. In this research, a Nd:YAG laser was utilized to produce 3136 Data Matrix symbols onto aluminum substrates. The experiment focuses on all interval values of the vector step, inter-step time, laser Q frequency and laser Q release time. From the experiment results, the optimized parameters are the vector step of 0.005~0.009 mm, the inter-step time of 29~43 μs, laser Q frequency of 7~10 KHz, and laser Q release time of 13~19 μs. And then, by analyzing the higher and the lower power density processing module SEM images and EDS data, it was found that the microstructure and micro-components of formations that created by the interaction between the laser and the aluminum alloy can affect the barcodes grade level significantly.