Unlike 201Tl, 99mTc-methoxy-isobutyl-isonitrile (99mTc-MIBI) does not redistribute in the myocardium after injection. Thus, two separate injections of this new myocardial perfusion agent are required to differentiate ischemia from scar. An injection at stress followed by a 2nd injection at rest performed 24 h later, or the inverse, has been proposed. This protocol is not ideal in clinical practice. It would be preferred if both injections were performed on the same day. Fifteen patients with significant coronary artery disease demonstrated by coronary angiography and with at least one ischemic segment on the myocardial 201Tl study were evaluated within two weeks with the following protocol. On the same day (short time interval protocol), 3 images, each of 10 min duration (anterior, 45 degrees LAO and 70 degrees LAO views) were performed between 30 to 60 min after the injection at rest of 7-10 mCi 99mTc-MIBI. On completion of the rest study, the patient received 25-30 mCi 99mTc-MIBI at stress and images were again obtained 30 to 60 min later. Two days later (long time interval protocol) a stress study alone was repeated using 10 mCi 99mTc-MIBI with the same imaging time. Qualitative and quantitative comparisons between the short and the long time interval studies were performed by four experienced observers. Both protocols showed the same number of ischemic segments (52/225) and fixed defects (19/225). The diagnostic information of images was judged similar in nine patients while the short protocol was judged superior to the long protocol in five patients and inferior in 1.(ABSTRACT TRUNCATED AT 250 WORDS)