To date, the architecture of supraspinatus (SP) and its relation to joint position has not been investigated. The purpose of this study was to quantify the dynamic architecture of the distinct regions of SP using ultrasound (US). Seventeen subjects (8 M/9 F), mean age 36.4 +/- 12.7 years, without tendon pathology were recruited. The SP was scanned in relaxed and contracted states. For the contracted state, SP was scanned with the shoulder actively abducted to 60 degrees and the glenohumeral joint in neutral rotation; 80 degrees external rotation; 80 degrees internal rotation. Fiber bundle length (FBL) and pennation angle (PA) of distinct regions, and muscle thickness were computed. Measurements of the posterior region were limited because of acromion shadowing. Parameters between regions and changes between relaxed and contracted states were analyzed using paired t-tests and repeated measures ANOVA (P < 0.05). On contraction in the anterior region, mean percentage of FBL shortening ranged between 9% and 21%. However, in the posterior region, shortening of approximately 2% only occurred in two of the three positions; lengthening of approximately 2.5% occurred in internal rotation. For the anterior region, the mean PA increased the least in the externally rotated position, and the mean PA of the middle part was smaller than the deep part for all states. Findings suggest changes in the architecture are not uniform throughout the muscle and joint position may play an important role in force production. The US protocol may serve as an outcome measure of adaptive changes of muscle function following surgery, training, and rehabilitation.