Laser based methods are well suited for automation and can be used to selectively remove dental caries to minimize the loss of healthy tissues and render the underlying enamel more resistant to acid dissolution. The purpose of this study was to determine which imaging methods are best suited for image-guided ablation of natural non-cavitated carious lesions on occlusal surfaces. Multiple caries imaging methods were compared including near-IR and visible reflectance and quantitative light fluorescence (QLF). In order for image-guided laser ablation to be feasible, chemical and physical modification of tooth surfaces due to laser irradiation cannot greatly reduce the contrast between sound and demineralized dental hard tissues. Sound and demineralized surfaces of 48 extracted human molar teeth with non-cavitated lesions were examined. Images were acquired before and after laser irradiation using visible and near-IR reflectance and QLF at several wavelengths. Polarization sensitive-optical coherence tomography was used to confirm that lesions were present. The highest contrast was attained at 1460-nm and 1500–1700-nm, wavelengths coincident with higher water absorption. The reflectance did not decrease significantly after laser irradiation for those wavelengths.