Effects of radiofrequency ablation (RFA) treatment of atrial fibrillation can be limited by the ability to characterize the tissue in contact. Parameters obtained by conventional catheters, such as impedance and temperature can be insufficient in providing physiological information pertaining to effective treatment. In this report, we present a near-infrared spectroscopy (NIRS)-integrated catheter capable of extracting tissue optical properties. Validation experiments were first performed in tissue phantoms with known optical properties. We then apply the technique for characterization of myocardial tissues in swine and human hearts, ex vivo. Additionally, we demonstrate the recovery of critical parameters relevant to RFA therapy including contact verification, and lesion transmurality. These findings support the application of NIRS for improved guidance in RFA therapeutic interventions.