Objective: To investigate the reliability of ultrasonographic measurement of acromiohumeral distance (AHD) and the effects of shoulder positioning on AHD among manual wheelchair users (MWUs) with spinal cord injury (SCI) and an able-bodied control group. Methods: Ten MWUs with SCI and 10 able-bodied subjects participated in this study. The ultrasonographic measurements of AHD from each subject were obtained by two raters during passive and active scapular plane arm elevation in neutral, 45°, 90°with and without resistance and in a weight relief raise position. The measurements were recorded again by each rater using the same procedures after a 30-minute time interval. All raters were blinded to each other's measurements. Setting: University Laboratories and Veteran Affairs Healthcare System. Results: Intra-rater (intraclass correlation coefficient, ICC > 0.83) and inter-rater (ICC > 0.78) reliability was excellent for both the MWUs with SCI and able-bodied groups across all arm positions except for the 45°p osition in the control group for one of the raters (intra-rater: ICC < 0.40 and inter-rater: ICC < 0.60). AHD significantly reduced when the shoulder was in the 90°arm elevated positions with or without resistance. Conclusion: Findings from our study demonstrated that ultrasonography is a reliable means to evaluate AHD in both able bodied and individuals with SCI, who are known to have significant shoulder pathology. This technique could be used to develop reference measures and to identify changes in AHD caused by interventions.