Background The aim of this study was to evaluate the value of multilayer strain analysis to the assessment of myocardial viability ( MV ) through the comparison of both speckle tracking echocardiography and single‐photon emission computed tomography ( SPECT ) imaging. We also intended to determine which segmental longitudinal strain ( LS ) cutoff value would be optimal to discriminate viable myocardium. Methods We included 47 patients (average age: 61 ± 11 years) referred to our cardiac imaging center for MV evaluation. All patients underwent transthoracic echocardiography with measures of LS , SPECT , and coronary angiography. Results In all, 799 segments were analyzed. We correlated myocardial tracer uptake by SPECT with sub‐endocardial, sub‐epicardial, and mid‐segmental LS values with r = .514 P < .0001, r = .501 P < .0001, and r = .520 P < .0001, respectively. The measurements of each layer strain (sub‐endocardial, sub‐epicardial, and mid) had the same performance to predict MV viability as defined by SPECT with areas under curve of 0.819 [0.778–0.861, P < .0001], 0.809 [0.764–0.854, P < .0001], and 0.817 [0.773–0.860, P < .0001], respectively. The receiver‐operating characteristic analysis yielded a cutoff value of −6.5% for mid‐segmental LS with a sensitivity of 76% and specificity of 76% to predict segmental MV as defined by SPECT . Conclusions Multilayer strain analysis does not evaluate MV with more accuracy than standard segmental LS analysis.