With advances in digitization, learning processes can be supported in many ways. The expectation that pre-service teachers learn to design, choose, and use appropriate digital multimedia in their prospective teaching also increases. A university course should thus help student teachers develop their intention and behavior for the future use of digital media in teaching by reinforcing their attitude, subjective norm, and self-efficacy. A crucial aspect when using digital media for teaching and learning purposes is to understand how learners perceive and learn with them. Assessing perceptual processes in multimedia learning with eye-tracking technologies can serve as a means to support student teachers in evaluating learners’ perceptions and cognitive processes. This might initiate a meaningful reflection about multimedia design and could positively influence student teachers’ behavioral intention for future use of digital media. We present a new course design that was implemented and explored with a cohort of advanced chemistry student teachers. Participants in this course (a) created learning materials respecting multimedia design principles, (b) investigated the learning processes with these materials in a small empirical eye-tracking study, and (c) reflected on purposeful design of teaching materials in scientific posters. The overall course design, evaluation results, and impact on student teachers’ attitude, subjective norm, and self-efficacy expectations toward the use of digital media in teaching are given. Implications for implementing multimedia learning with eye-tracking in a student teacher course are also discussed.