The goal of this study was to visualize the supraspinatus tendon three-dimensionally using fast spin-echo (FSE) MRI and validate the accuracy of measuring the dimensions of the supraspinatus tendon based on 3D reconstructed images. Nine cadaver shoulders (51–84 y/o, mean 70.0 y/o) were imaged at conventional T2-weighted spin-echo (CSE), gradient echo (GRE), and 3D T2-weighted FSE sequences. Each "object" of the supraspinatus muscle, tendon and scapula was three-dimensionally reconstructed using ANALYZE™ image data processing software. The FSE images revealed significantly higher contrast of the tendon and contrast-to-noise ratios of the fat-to-tendon and fat-to-muscle. The length of the anterior, middle, and posterior portions of the tendon were measured in two ways: (1) from the three-dimensional reconstructed images, and (2) directly from the cadaver specimen using calipers. No statistically significant differences were found between the ANALYZE™ and caliper measurements using a paired t-test for the anterior (p = 0.55), middle (p = 0.57) and posterior (p = 0.44) portions of the supraspinatus. The 3D FSE sequence exhibits higher spatial resolution, spends shorter acquisition time, and constructs a voxel data set. These advantages can prevent blurring artifacts when imaging the supraspinatus tendon of a human body. Tendon length measurements derived from three-dimensional reconstructions using ANALYZE™ were found to be good estimates of actual tendon length. Therefore, the combination of FSE sequence and 3D image data processing provides a method for noninvasive quantitative analysis of supraspinatus tendon morphology. The results lay the groundwork for future quantitative studies of cuff pathology.