The aim of the present study was to investigate the cariostatic effect of titanium tetrafluoride (TiF4) on in situ-induced lesions on human root surfaces. Analysis of the samples was carried out by quantitative microradiography (TMR) and confocal laser scanning microscopy (CLSM). Pre-molar roots, sectioned into four pieces, were used for sample preparation. Before they were mounted into the recesses prepared in the acrylic intra-oral appliances, two root pieces from each tooth were treated with 4% TiF4 for 1 min, while the remaining two pieces served as controls. The appliances were worn by 12 volunteers for 4 wks. After 4 wks with no fluoride supplementation, the root pieces were removed and analyzed by TMR and CLSM. The TMR results showed that the TiF4 treatment reduced lesion depth and total mineral loss by 56% and 62%, respectively. CLSM images agreed well with the TMR measurements. A dense light-reflecting surface layer with almost intact subsurface structures was evident in the TiF4-treated samples.