This study investigated the utility of optional thallium-201 (201Tl) imaging for detecting myocardial viability in the stress 99Tcm-tetrofosmin/rest 201Tl dual-isotope protocol. Seventy-nine patients with old myocardial infarction and 25 patients with acute myocardial infarction underwent acquisition of three consecutive 201Tl images (early, intermediate and delayed) using the dual-isotope protocol. A polar map was created and defect scores (extent and severity) were determined by comparison with normal control data. Fluorodeoxyglucose positron emission tomography was also performed in 16 patients with old myocardial infarction. In patients with old infarction, the severity score decreased significantly from the early to the intermediate images, and decreased further on the delayed images. In patients with acute infarction, the score increased from the early to the intermediate images, but not on the delayed images. Regional uptake on the delayed images showed a better correlation with the fluorodeoxyglucose images than that on the early images. Redistribution on the delayed images was exclusively observed in the myocardial segments with less uptake than that estimated by fluorodeoxyglucose. In conclusion, addition of delayed 201Tl imaging to the dual-isotope protocol could improve the sensitivity for detecting myocardial viability.