Autofluorescence Loss in Photobleaching for Human Dentin ex vivo

Two-photon fluorescence microscopy was performed on the enamel-dentin junction area of a human tooth using a femtosecond pulsed laser. We obtained a clear image contrast between the bright dentin and dark tubules with the autofluorescence generated from the endogenous fluorophores in dentin. The autofluorescence shows a broad spectrum due to complex cross links between dentinal collagens, which extend from blue to orange wavelengths (470???590 nm), but a gradual autofluorescence loss in photobleaching was observed for a long-term exposure under strong excitation. For increasing excitation power, we found that two-step decay becomes significant in the spectrally integrated autofluorescence.