Image subtraction techniques, in conjunction with [75Se]selenomethionine (75Se) scintigraphy, have recently been suggested to be a potentially valuable tool for localization of parathyroid abnormalities. With use of these techniques, we prospectively studied 15 patients scheduled for parathyroid operations. Postoperatively, all were normocalcemic. Anterior scintiscans of the neck were divided into quadrants, and regions of enhanced uptake were assigned to one or more quadrants. On this basis, by chance alone a minimum of 25% of single-gland enlargements would be assigned to the correct quadrant of the neck. We found that by use of 75Se scanning only 8 of 22 abnormal glands (36%) were assigned to the quadrant of the neck in which they were found intraoperatively. In no case was a clearly visualized focus of parathyroid activity encountered. These results do not justify the continued use of 75Se scintigraphy with or without image subtraction in the preoperative localization of parathyroid glands.