Catheter-based radiofrequency ablation for atrial fibrillation has long-term success in 60-70% of cases. A better assessment of lesion quality, depth, and continuity could improve the procedure’s outcome. We investigate here photoacoustic contrast between ablated and healthy atrial-wall tissue in vitro in wavelengths spanning from 410 nm to 1000 nm. We studied single-and multi-wavelength imaging of ablation lesions and we demonstrate that a two-wavelength technique yields precise detection of lesions, achieving a diagnostic accuracy of 97%. We compare this with a best single-wavelength (640 nm) analysis that correctly identifies 82% of lesions. We discuss the origin of relevant spectroscopic features and perspectives for translation to clinical imaging.