Metacognition is the ability to monitor and evaluate one's thoughts about learning and has been shown in some studies to improve the effectiveness of instructors. With dissection-based gross anatomy, instructors dynamically use their metacognition to monitor student learning and adapt their teaching at tableside. This study explored the metacognition of instructors as they taught in an anatomy course for allied health students. All instructors in a doctoral-level gross anatomy course at a single institution, including faculty, associate instructors (AIs), and teaching assistants (TAs), were invited to participate. At the start and end of the course, participating instructors completed pre- and post-questionnaires, which included the Teacher Metacognition Inventory (TMI), a 28-item survey that assesses metacognition as it relates to teaching. After labs, instructors completed reflective journals to provide deeper insight into their metacognition. Reflective journals were then thematically analyzed. Thirteen (52%) instructors participated in this study, including five faculty, three AI, and five TAs. Between the start and end of the course, total TMI score increased from 108.8 to 114.3 (p = 0.046). TAs exhibited the greatest change in total TMI score ([delta]mean = +11.4) followed by faculty ([delta]mean = +3.2) and AIs ([delta]mean = -0.7). Several themes were also identified. For example, TAs were more internally focused on content mastery, whereas faculty were externally focused on interpersonal factors (e.g., inclusive language). These insights into the metacognition of anatomy instructors, though limited in reliability and generalizability, may inform how to best support their professional development. Novices may benefit from content reviews, while experienced instructors may benefit from inclusivity or communications training.