The objective of this study was to analyse the effectiveness of some parameters which characterise the change in morphology in human root canals subjected to ProTaper rotary enlargement with the help of an X-ray microfocus computed tomography (MCT) and to introduce a novel parameter that is effective in quantifying changes in root canal morphology. Ten each straight and curved root canals with mature apices chosen from extracted human upper incisor and canine teeth were scanned with MCT before and after canal shaping using ProTaper rotary instruments in order to facilitate three-dimensional digital reconstruction and quantitative gauging of relevant instrumental parameters and changes therein (surface area and volume). Root canal geometry change and the effectiveness of shaping were quantified with Structure Model Index change (Delta SMI) and surface area change to volume change ratio (Delta SA/Delta V). These two parameters were also tested on simulated canals. Postinstrumentation cross-sectional changes were also analysed, but only on the plastic blocks. Statistical analysis of parameters was carried out to verify the significance of results. Analysis of cross-sectional shape of postinstrumented resin simulated canals showed statistically significant decrease in Form Factor (p<0.05) and statistically significant increase in Eccentricity (p<0.005). Delta SMI did not show significant difference between straight and curved canals. SMI values showed bidirectional change during root enlargement which questions the reliability of this metric in analysing instrumentation. Statistically significant (p<0.005) deviations in Delta SA/Delta V were quantified as 1.92 and 3.22 for straight and curved human canals, respectively. Instrumentation-induced canal geometry change was determined to be more pronounced in curved canals using the novel parameter Delta SA/Delta V. This has been proven as being a statistically accurate and reproducible parameter for quantitative characterisation of root canal geometry change and differentiation of preparational efficacy for both straight and curved root canals.