Simultaneous investigations with two isotopes are currently restricted because of spectral overlap. The factor analysis of spectral and dynamic structures (FASDS) method is shown to achieve accurate spectral separation. In addition, it estimates underlying dynamic mechanisms. Twenty-six patients were injected simultaneously with 99Tcm-pertechnetate and 201Tl-chloride to assess the malignancy of solitary thyroid nodules. List-mode acquisition of spectral, temporal and spatial coordinates of events allows the reconstruction of an image sequence indexed by time and energy. FASDS proceeds in two steps. First it yields both dynamic and spatial information related to each isotope (99Tcm and 201Tl) and partially removes the scatter component. Then it estimates the underlying kinetics and associated spatial distributions of each isotope. Using the 201Tl component, an index was derived from the uptake ratio between nodules and normal thyroid tissue. Concerning the detection of malignant nodules the method indicated no false negative in our limited group of 26 patients. One false positive result was found which could not be classified by the investigation of the 201Tl dynamic components contained in the reconstructed 201Tl factor sequence.