The objectives of this study were to determine systematic changes of the normal subacromial space width during abduction and rotation, and to analyze the spatial relationship of the supraspinatus muscle with the acromion and clavicle. 12 healthy volunteers were imaged by an open MR scanner in 5 different positions of abduction and in 3 positions of rotation. After three dimensional (3D) reconstruction and 3D Euclidian distance transformation, the minimal spatial distances between the humerus and the acromion and the humerus and clavicle were computed. The minimal acromio-humeral distance decreased significantly from 30 degrees of abduction (mean 7.0 mm +/- 1.6 mm) to 120 degrees (mean 3.9 mm; +/- 1.8 mm; p < 0.0001). At 30 degrees, the minimal distance penetrated the supraspinatus, whereas at 120 degrees it was always located lateral to the supraspinatus tendon. At 90 degrees with internal rotation (7.6 mm, +/- 2.3 mm) the minimal acromio-humeral distance was larger than in neutral rotation (5.4 mm, +/- 2.3 mm) or external rotation (4.4 mm, +/- 2.2 mm; p < 0.05), but it penetrated the supraspinatus tendon at its most vulnerable part, reaching the acromion at its anterior inferior border. We conclude that the subacromial space width changes during abduction and rotation and that the supraspinatus is in closest contact to the anterior inferior border of the acromion in 90 degrees of abduction with 45 degrees internal rotation. These values obtained in volunteers can be used as a basis for further investigations in patients with the impingement syndrome.