Background: Early childhood educators (ECEs) influence young children’s early uptake of positive health behaviours in childcare settings and serve as important daytime role models. As such, it is imperative that post-secondary early childhood education programs provide students with the foundational knowledge and professional training required to confidently facilitate quality active play opportunities for young children. The primary objective of the Training pre-service EArly CHildhood educators in physical activity (TEACH) study is to develop and implement an e-Learning course in physical activity and sedentary behaviour to facilitate improvements in: pre-service ECEs’ self-efficacy and knowledge to lead physical activity and outdoor play opportunities and minimize sedentary behaviours in childcare. This study will also explore pre-service ECEs’ behavioural intention and perceived control to promote physical activity and outdoor play, and minimize sedentary behaviour in childcare, and the implementation of the e-Learning course. Methods/Design: A mixed-methods quasi-experimental design with three data collection time points (baseline, post-course completion, 3-month follow-up) will be employed to test the e-Learning course in early childhood education programs (n = 18; 9 experimental, 9 comparison) across Canada. Pre-service ECEs enrolled in colleges/universities assigned to the experimental group will be required to complete a 4-module e-Learning course, while programs in the comparison group will maintain their typical curriculum. Pre-service ECEs’ self-efficacy, knowledge, as well as behavioural intention and perceived behavioural control will be assessed via online surveys and module completion rates will be documented using website metrics. Group differences across timepoints will be assessed using linear mixed effects modelling and common themes will be identified through thematic analysis. Discussion: The TEACH study represents a novel, evidence-informed approach to address the existing gap in physical activity and sedentary behaviour-related education in Canadian post-secondary early childhood education programs. Moreover, e-Learning platforms, can be employed as an innovative, standardized, and scalable way to provide ECEs with consistent training across jurisdictions.