To promote the cultivation of undergraduates’ scientific research innovative thinking and practical ability, and to practice the collaborative education mechanism of “research and teaching”, the Langmuir-Blodgett (L-B) self-assembly technology to prepare a highly ordered polystyrene (PS) microspheres array experiment is taken as an example, which requires students to analyze the morphology, degree of order and laser 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. It is beneficial for improving student’s scientific consciousness and practical ability, and it provides an advantageous exploration for cultivating new talents with innovative scientific thinking.