Teachers' professional vision (TPV) is an essential contributor to enhancing the quality of teaching-learning processes thanks to its potential effect on teachers' self-reflective skills. This is of particular importance in Physical Education (PE), due to the fact that this subject presents different challenges and more dynamic situations compared to others. One of the latest approaches for enhancing TPV is video-based reflection, allowing teachers to observe and analyze recorded lessons. This research aimed to test the effect of a video-based reflection program on TPV in PE. Two teachers participated in a 3-step video-analysis program consisting of self-reflection, peer reflection, and expert feedback. Notable changes were found in TPV at the end of the 6-month intervention, both in terms of teachers' selective attention to classroom events and their knowledge-based reasoning. Our findings suggest that video-based reflection interventions could represent an important component of any teacher training program aiming to increase PE teachers' ability to evaluate and respond to a variety of in-class situations.