The autopsy and surgical pathology databases at UCLA were searched for cases of TOF and its major variants (including TOF with pulmonary atresia and TOF with absent pulmonary valve). Baseline demographic and clinical data were obtained from the medical records. All specimens were evaluated for the presence of naturally or surgically related anatomic isthmuses corresponding to those that have been previously described. In addition, isthmus 1 was redesignated as 1A, 12 and © 2013 American Heart Association, Inc. Original ArticleBackground-Although catheter ablation has been used to target the critical isthmuses for re-entrant monomorphic ventricular tachycardia in tetralogy of Fallot, the anatomy and histology of these regions have not been fully characterized. Autopsy hearts with tetralogy of Fallot were evaluated to clarify the pathological substrate. Methods and Results-Twenty-seven hearts with the diagnosis of tetralogy of Fallot were examined. Anatomically defined isthmuses included (1A) ventriculotomy-to-tricuspid annulus, (1B) ventriculotomy-to-ventricular septal defect patch, (2) ventriculotomy-to-pulmonary annulus, (3) pulmonary annulus-to-ventricular septal defect patch, and (4) ventricular septal defect patch-to-tricuspid annulus. Length and wall thickness were measured for all specimens, and light microscopy was performed for those surviving surgery. For subjects ≥5 years at death, isthmuses 1A and 1B were present in 88%, isthmus 2 in 25%, isthmus 3 in 94%, and isthmus 4 in 13%. Isthmus 1A had the greatest dimensions (mean length, 3.9±1.08; thickness, 1.5±0.3 cm), isthmus 1B intermediate dimensions (mean length, 2.4±0.8; thickness, 1.1±0.4 cm), and isthmuses 2, 3, and 4 the smallest dimensions (mean length, 1.5±0.5, 1.4±0.8, and 0.6±0.4 cm; thickness, 0.5±0.2, 0.6±0.2, and 0.3±0.04 cm, respectively). Histological examination (n=7) revealed increased fibrosis in anatomic isthmuses relative to nonisthmus controls. Conclusions-Consistencies in isthmus dimensions and histology are found among patients with repaired tetralogy of Fallot. Isthmus 1A is associated with the largest morphological dimensions, whereas the nearby newly described isthmus 1B is significantly smaller. Of isthmuses with the smallest dimensions, isthmus 3 is the most common. (Circ Arrhythm Electrophysiol. 2013;6:905-911.)