Abstract: To promote the cultivation of undergraduates’ scientific research innovative thinking and practical ability, and to practice the collaborative education mechanism of “research and teaching”, and by taking the Langmuir-Blodgett self-assembly technology to prepare a highly ordered polystyrene (PS) microsphere array experiment as an example, the experiment is required to analyze the morphology, degree of order and optical diffraction of the as-prepared samples. This comprehensive experiment covers multiple knowledge points such as the principle of self-assembly, structural fabrication, morphology characterization and two-dimensional laser diffraction, etc. It is beneficial for improving student’s scientific consciousness and practical ability, and makes a beneficial exploration for cultivating new talents with innovative scientific thinking.