Prior research has explored various factors to explain differences in teaching experiences and behaviors among school teachers, including self-efficacy, basic psychological need satisfaction, and emotions. However, these factors have predominantly been examined in isolation, and limited research has investigated their role in the context of higher education teaching. To address these research gaps, analyses on both the within and between teacher level are needed. The aim of the present study was therefore to investigate the interplay between these motivational and emotional constructs on both levels, as well as the relevance and applicability of prior research findings on school teachers to the context of higher education teaching at universities. In a micro-longitudinal study, 103 university teachers from Germany (49 female; average age: 41.4 years, SD = 11.0) completed assessments of their self-efficacy in 748 sessions directly before their teaching sessions, as well as their basic psychological need satisfaction and discrete emotions directly after. Multilevel structural equation modeling revealed positive associations between self-efficacy and basic psychological need satisfaction. Self-efficacy was negatively associated with negative emotions, and positive indirect effects on positive emotions as well as negative indirect effects on negative emotions were identified through satisfaction of the needs for competence and relatedness. Basic psychological need satisfaction was positively related to positive emotions and vice versa--however, unexpected positive associations between relatedness and negative emotions emerged and require further research.