Purpose/Hypothesis: The profession of physical therapy has undergone an evolution since its inception, growing from a technical training program to a doctoral program. Anatomy is considered a foundational science for the physical therapy profession and is an integral part of the human movement system. Despite a breadth of research, little consensus has been reached regarding the most efficient method of anatomy instruction in regards to student grades or anatomy knowledge of physical therapists. The objective of this study was to determine knowledge outcomes based on the anatomy instructional method provided in physical therapist educational programs. Specifically, the hypothesis investigated was that physical therapists who were provided anatomy lecture and laboratory educational exposure in physical therapy school would show a higher level of anatomy knowledge. Number of Subjects: 3,535 Materials and Methods: : The 3,535 participants, 65.7% female, 33.3% male, and 0.1% other, completed a questionnaire demonstrating their knowledge of anatomy topics in six (6) regional areas (subsets): 1) upper extremity; 2) lower extremity; 3) thorax and abdomen; 4) pelvis; 5) spine; and 6) head. For each regional area, participants provided the method of anatomy instruction they received in physical therapy school. Instructional methods included lecture combined with one of the following laboratory instructional methods: 1) dissection; 2) prosection; 3) plastination; 4) technology driven; 5) no laboratory; and 6) no lecture or laboratory instruction. Results: Instructional method used in physical therapy school showed significant differences in two regional anatomy subsets scores. 1) the dissection laboratory method (n= 2,879, M=2.52, SD= 1.05) exhibited higher means when compared to the prosection laboratory method (n= 440, M=2.40, SD=1.08) in the thoracic subset; 2) the dissection laboratory method (n= 2,288, M=2.41, SD= 1.22) exhibited higher means when compared to the no laboratory method (n= 199, M=2.00, SD=1.14) and when compared to the prosection laboratory method (n=769, M= 2.20, SD=1.23) in the head subset. Conclusions: Findings were consistent with the hypothesis that comprehensive exposure to the thoracic and head regional subsets lead to greater demonstration of anatomy knowledge. No significant knowledge differences were seen in the four other regions. The variance explained in anatomy knowledge based on instructional method was small, 0.4%, indicating other factors play a role in the anatomy knowledge of physical therapists. Clinical Relevance: This study investigated how anatomy education methods within physical therapist educational programs play a role in anatomy knowledge of physical therapists. In current physical therapist practice, there exist practitioners with varying clinical degrees, backgrounds of didactic curricula, continuing professional education experiences, and self-defined expertise. Despite these differences, this study provided evidence supporting the use of dissection laboratory instruction in the thoracic and head regions.