Raman spectroscopy (RS) has emerged as a method of choice for label-free, nondestructive, objective, and rapid analysis of biological samples. RS has found profound applications in dental research with studies on early caries detection, carbonate and phosphate contents, mineralization and demineralization patterns and stresssignatures in teeth samples. Raman dental spectral features are usually affected by fluorescence, especially archaeological samples, and are prone to thermal damage when laser power is increased for superior signal-to-noise ratio. This affects the quality of Raman dental maps, limiting their utility. This also warrants the need for exhaustive spectral preprocessing. One way to refine signal quality is through enhanced collection efficiency. In this study, we explored if spectra-quality from dental samples can be improved by signal collection through water-immersion objectives, since informative Raman regions are not affected by water. Raman maps were obtained from tooth samples under air-immersion objectives (10X and 50X) and a water-immersion objective (63X) using 532 nm laser. Spectra acquired through the different objectives were compared. We observed that the water-immersion objective reduced fluorescence, decreased baseline, and prevented thermal heating/damage to the samples. Dental samples were not affected by prolonged immersion in water. The improved spectral quality allows lower acquisition-time for mapping experiments. Further, it also reduces the preprocessing required to obtain sufficient signal-to-noise quality. Thus, application of water-immersion objectives in Raman experiments has the potential to enhance spectral quality and experimental outcomes while keeping the samples preserved for further analyses.
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