Background: Atrial tachycardia (AT) is a commonly encountered rhythm disorder in patients with underlying atrial scar. The role of atrial late activation mapping during sinus rhythm to predict the critical isthmus (CI) of AT has yet to be systematically evaluated. We aimed to investigate the relationship between the functional substrate mapping (FSM) characteristics and the CI of reentrant ATs in patients with underlying atrial low-voltage areas. Methods: Patients with history of left AT who underwent catheter ablation with 3D mapping using high-density mapping were enrolled. Voltage map and isochronal late activation mapping were created during sinus/paced rhythm to detect deceleration zones (DZ). Electrograms with continuous-fragmented morphology were also tagged. After induction of AT, activation mapping was performed to detect CI of the tachycardia. Atrial tachyarrhythmia (ATa) recurrence was defined as detection of atrial fibrillation or AT (≥30 s) during the follow-up. Results: Among 35 patients [mean age: 62 ± 9, gender: 25 (71.5%) female] with left AT, a total of 42 reentrant ATs induced. Voltage mapping during sinus rhythm revealed low-voltage area of 37.1 ± 23.8% of the left atrium. The mean value of bipolar voltage, EGM duration, and conduction velocity during sinus rhythm corresponding to CI of ATs were 0.18 ± 0.12 mV, 133 ± 47 ms, and 0.12 ± 0.09 m/s, respectively. Total number of DZs per chamber was 1.5 ± 0.6, which were located in the low-voltage zone (<0.5 mV) detected by high-density mapping. All CIs of reentry were colocalized with DZs detected during FSM. The positive predictive value of DZs to detect CI of inducible ATs is 80.4%. Freedom from ATa after the index procedure was 74.3% during a mean follow-up of 12.2 ± 7.5 months. Conclusion: Our findings demonstrated the utility of FSM during sinus rhythm to predict the CI of AT. DZs displayed continuous-fragmented signal morphology with slow conduction which may guide to tailor ablation strategy in case of underlying atrial scar.