Visualization of initial bacterial colonization on dentin using fluorescence activating headlight for fluorescence enhanced theragnosis

•Accuracy of caries detection and the application-sensitivity of the new designs for vision's ground-breaking REVEALTM utilizing a fluorescence activating headlight for excitation purpose.•Excitation of electrons from the ground state energy level to a higher vibrational energy level causes fluorescent and bioluminescent light emission.•Benefits to employing bioluminescence instead of fluorescence as a diagnostic tool in dentistry. To evaluate accuracy of caries detection and the application-sensitivity of the new Designs for Vision's REVEAL™ utilizing a fluorescence activating headlight for excitation purpose. REVEAL dental fluorescence loupes and headlight system were used. Occlusal enamel was removed, and mid-coronal dentine was exposed. Carious artificial lesion was created. Streptococcus mutans, Actinomyces naeslundii, and Streptococcus sanguis were used. The assessment was performed using two diagnostic methods: naked eye and Design for Vision Glasses with inter examiner blinding using two calibrated examiners. After 7 days, Raman measurements were made on dentin disc specimens with 785 nm wavelength. The bacterial counts in colony-forming units (CFU) were used to examine the growth kinetics of biofilms. The collagen fibril structure within the discs was performed using Transmission Electron Microscope. Scanning Electron Microscope was used to image samples at various magnifications. FISH was performed with specimens fixed in 4% paraformaldehyde in phosphate-buffered saline. Reproducibility was measured by Cohen kappa scores, values of which range from 0 for less than chance agreement to 1 for almost perfect agreement (p