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This mixed-methods study investigated how preservice elementary teachers’ (n=465) integrated STEM (iSTEM) teaching self-efficacy and perceived challenges shift during their semester-long methods coursework. Quantitative data sources included pre-post measures using the Self-Efficacy for Teaching Integrated STEM instrument, an open-ended questionnaire, and demographic questions. Quantitative analysis using McNemar and t-tests showed statistically significant positive gains in iSTEM teaching self-efficacy and a significant decrease in overall perceived challenges by the end of the semester. Qualitative content analysis revealed six categories of perceived challenges: (1) teacher affect and experience, (2) student-related concerns, (3) content and curricular challenges, (4) pedagogical challenges, (5) support from others, and (6) time and resources. Implications for STEM teacher preparation programs and future research will be discussed.