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Electron probe microanalysis of permanent human enamel and dentine. A methodological and quantitative study.

Sample preparation of dental tissues for quantitative electron microprobe analysis has not been critically examined because of the highly mineralized nature of these structures. The present study was designed to establish the most suitable method for the electron probe quantitative determination of calcium in human permanent enamel and dentine while preserving the morphological features. Comparisons of quantitative data obtained with air-drying and freeze-drying methods showed that calcium in enamel was more accurately measured in specimens prepared with cryopreservation and freeze-drying. No significant differences between the methods tested were found in dentine although cryopreservation and freeze-drying yielded less statistical variability. Moreover this approach did not modify morphological features of interest. We recommend this combination of processing techniques for human permanent teeth not only because it was found the most accurate and least variable in determining calcium concentration, but also because of its potential usefulness in studies of alterations in tooth mineralization.