Rest technetium 99m-sestamibi uptake may underestimate myocardial viability in asynergic territories. Because nitrate administration was reported to improve thallium 201 uptake in perfusion defects, this study aimed to test the influence of nitrates on 99mTc-sestamibi uptake and on the tracer capability to recognize viable tissue in asynergic segments. In 23 patients with prior infarction and left ventricular dysfunction, regional wall motion was assessed by echocardiography before and after revascularization (13 segments/patient). Group 1 included 97 normokinetic; group 2, 97 hypokinetic; and group 3, 105 akynetic or dyskinetic segments; group 3 was divided into group 3A (72 segments unchanged after revascularization) and group 3B (33 segments with functional recovery). 99mTc-sestamibi uptake was graded using a scoring scheme in the same 13 segments both at rest and, on a separate day, injecting the tracer during isosorbide dinitrate infusion (ISDN). At rest, the mean 99mTc-sestamibi uptake decreased significantly from group 1 through group 3. With ISDN, the mean 99mTc-sestamibi uptake increased in all groups compared with rest, but the increase was significant only in groups 2 and 3, and within the latter, only in group 3B. Thus, with ISDN group 3B was no longer different from group 2. Only 6% of group 3A segments showed an improved uptake with ISDN, versus 33% of group 3B (P < .00005). At rest only 14 of 33 segments of group 3B showed a normal or slightly reduced uptake, whereas these were 25 of 33 with ISDN (P < .02). In conclusion, the acute administration of ISDN increases the uptake of 99mTc-sestamibi mainly in those asynergic territories that show postrevascularization functional recovery. Therefore, ISDN 99mTc-sestamibi imaging might improve the tracer capability to detect viable hibernating myocardium.