This work aims to explore whether students' learning outcomes in the virtual museum (VM) 's "Life and Science course" are influenced by the type of knowledge and the different levels of interaction within the museum. The study was conducted using an eye-movement experimental method, based on cognitive load theory, knowledge visualisation and connectionist theory, to construct a virtual museum interaction hierarchy index to identify high and low levels of interaction in the experimental material. A randomised two-factor experiment was organised to obtain eye-movement related indicators and to analyse statistical data in conjunction with retention test scores and transfer test scores, taking into account the influence of the perceived difficulty of the learning material and the level of mental effort of the students on the experiment. This study will provide effective suggestions for museum learning design and resource development, ultimately offer insights in exploring the most suitable learning rules and learning material selection for VM learners.