In order to improve the fabricating efficiency ofn micro-and nano-structures by laser-induced backward transfer technology, a three-beams laser-interference-induced backward transfer (LIIBT) method was established to fabricate micro-nano structure arrays, which was based on laser interference and laser-induced backward transfer technology. In this article, Indium Tin Oxide (ITO) glass was used as a substrate to receive an Au nanoparticles jet from Au thin film. The LIIBT was carried out by three-beams laser interference. The Scanning Electron Microscopy(SEM) images indicate that the a perfect 5-μm period structure could be fabricated under the laser fluence of 25 mJ/cm2 for the Au 50 nm thin film 50 nm. Meanwhile, uniform distribution of the Au nanoparticles could be observed on the surface of the period structures, and the over 80% of Au nanoparticles over 80% were smaller than 100 nm. The Energy-Dispersive X-ray spectroscopy (EDX) analysis shows that the microscale period structures were composed of a lot of Inand Au elements, which prove validates the fabrication of theisemicroscale period structures due to the laser pulse ablation on the ITO layer, and then, Au nanoparticles were deposited on it, simultaneously. Finally, the SERS was studied by the Raman spectrum for the Au nanostructures after coating them with 1.0×10-5, 1.0×10-7, and 1.0×10-9 M rhodamine 6G. The results show that these peaks are located at 612 cm-1, 773 cm-1, 1 190 cm-1, 1 319 cm-1, and 1 511 cm-1, which indicate that the Au nanostructures had obvious SERS effect for the trace of the R6G. The LIIBT will be used to improve the efficiency of the laser-induced backward transfer technique on the micro and nanostructure. This technique, which will have the potential applications for the ultra-sensitive detection, optoelectronic devices, and microfluidic flow control systems.