To correlate dental age with an individual's chronological age based on the calculated volume ratio of pulp versus tooth volume measured, an X-ray microfocus computed tomography unit (μCT) with 25μm spatial resolution was used to non-destructively scan 43 extracted single root teeth of 25 individuals with well-known chronological age. Custom-made analysis software was used by two examiners to obtain numerical values for pulpal and tooth volume. The ratio of both was calculated and statistically processed. No significant intra-orinter-examiner differences were found. In fact,a very strong concordance correlation coefficient was found. Linear regression analysis showed a coefficient of determination (r) of 0.31 which suggests that there is a rather weak correlation between the volume ratio of pulp versus tooth and biological age. Although rather time consuming, this technique shows promising results for dental age estimation in a non-destructive manner using X-ray microfocus computed tomography.