The Four-Dimensional Ecology Education (4DEE) Framework was adopted by the Ecological Society of America as a way to enhance ecological literacy among college students. The framework encompasses Ecological Concepts (EC), Ecological Practice (EP), Cross-Cutting Themes (CCT), and the Human-Environment Interaction (HEI) dimensions. As the 4DEE framework is being adopted in more science classrooms, there is an increasing interest in assessing its effectiveness in achieving ecological literacy. This study aimed to (1) utilize the 4DEE framework to design an assessment tool for three courses, (2) develop a collaborative effort for assessment, and (3) explore how students engage with the multidimensional assessment. Students across three universities engaged in an interactive 3-dimensional lesson that focused on species interactions (EC), data analysis (EP) and ecosystem services, anthropogenic impacts on bee diversity (HEI). Overall, the results showed that students had a difficult time producing answers that incorporated multiple dimensions and may need more practice in the classroom. Across institutions most students enjoyed the module. Seventy-three percent of students mentioned they enjoyed the module, found it engaging and/or found it educational. As a whole, these data show that utilizing the 4DEE framework demonstrated students' understanding of ecological concepts and most students found it enjoyable.