Vital tooth bleaching can be achieved through the application of external sources of hydrogen peroxide derived from paint on, tray or strip applications. The verification of safety of novel bleaching formulations demands study of internal as well as superficial external effects of bleaching gels on tooth structures. Objectives: This study examined the effects of Crest Whitestrips Supreme (CWSS) bleaching strips containing 14 % hydrogen peroxide on surface and subsurface ultrastructural and microchemical properties of enamel and coronal dentin. Methods: Human molars were mounted in methacrylate blocks. Color determinations (CIELAB) of standardized areas on surface enamel were taken by image analysis. Teeth were bleached with CWSS for 42 hours of treatment in a cycling regimen including continuous pooled saliva immersion with bid ½ hour treatments in peroxide gels. A control group was untreated and cycled through saliva soaks. Following bleaching, color values were re-determined. Subsurface CLSM-microtomographic images (reflection and fluorescence mode) combined with micro-Raman spectra uptake (633 nm laser for reflection CLSM and Raman, 488 nm for fluorescence) were recorded on transverse sectioned teeth permitting subsurface evaluation of ultrastructural and microchemical properties. Results: Micro-Raman spectra were compared for test and control specimens as subsurface line scans in defined intervals. Raman bands associated with mineral PO4 (954 cm-1), CO3 (1071 cm-1) and non-apatite HPO4 (1002 cm-1) vibrations were evaluated to assess hard tissue integrity. The CLSM tomographies and Raman spectra of bleached enamel were unchanged from control enamel specimens. CWSS peroxide treatment, however, produced changes in autofluorescence of enamel and coronal dentin. Conclusions: CLSM tomographies and Raman spectra of enamel and dentin appear unchanged in the present bleaching process, being evidence for unaffected tissue ulta-structure. Decrease of auto-fluorescence and Raman background may be correlated with the elimination of non-functional chromophores.