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Diagrams are a visual representation found in nearly every science classroom. They are often critical for learning and can communicate information more efficiently than language alone. Despite the prevalence of diagrams, students often have difficulty understanding them. This has led to research evaluating how we can improve student diagram comprehension. A major barrier in this process is the lack of a robust measure of diagram comprehension that spans multiple scientific domains. The present study works on the development of the Science Diagram Comprehension Test (SDCT) and provides evidence of the SDCT’s reliability, content validity, internal structure validity, and criterion validity. The results suggest that the SDCT can serve as an effective measure for evaluating diagram comprehension interventions.