Abstract: The single-crystal X-ray diffraction experiment is a crucial instrumental analysis course in majors such as materials science, chemistry, and biomedical science. However, due to limited class hours, students struggle to fully master experimental skills under the traditional teaching mode. This study constructs an online-offline interactive hybrid teaching model for the experiment, covering pre-class online theoretical preview and quizzes, in-class guidance and experimental implementation, and post-class consolidation and extended practical training. The model aims to comprehensively enhance students’ skills in single-crystal X-ray diffraction experiments. Additionally, digital instructional resources for crystal structure determination and refinement, developed based on the Cambridge Structural Database (CSD), are introduced into the teaching process to address the current weakness in single-crystal structure determination and refinement in experimental instruction. Practical results indicate that the online-offline hybrid teaching model is highly effective, significantly improving students’ experimental proficiency and their skills in single-crystal structure determination and refinement.