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In higher education, academic success and failure can be affected by students' affective states, such as their attitudes towards courses and learning. Many studies highlight the role of attitudes in the educational process resulting in a high demand for attitude assessments to plan comprehensive instructional strategies. The present study assessed and compared attitudes towards science in higher education. A total of 296 Indonesian undergraduate students from science teacher training and engineering majors participated in this study by completing the online Attitudes Towards Science Questionnaire (ATSQ). The validation analysis with confirmatory factor analysis and the Rasch model was performed to measure the instrument's properties resulting in a good fit model for the questionnaire's theoretical construct with acceptable individual fit items. Comparing the attitudes of science teacher candidates and engineering students, no significant differences were found in attitude towards science. Both groups revealed moderate attitudes, with higher responses to the science value variable, followed by enjoyment and confidence, anxiety and difficulty, and participation in science learning and activities. However, there were differences in the pattern of correlations between the variables, especially for anxiety and difficulty. The ATSQ has good psychometric properties and therefore can be used in research to assess attitudes towards science. It can stimulate further research in different contexts to confirm empirical evidence and promote attitude development in higher education.