Aim: The present work aims to compare a digital dental histoanatomical analysis method with chemical enamel dissolution for the study of dental morphology. Material and methods: Extracted maxillary anterior teeth were scanned under microcomputed tomography (μCT) (μCT 40; Scanco Medical), segmented, and reconstructed three-dimensionally (Amira, version 5.5.2; VSG). Following the digital acquisition of dental morphology, all specimens were acid treated with 5% formic acid for careful dissolution of the enamel layer. Six measurements (three buccopalatal and three mesiodistal) per specimen were performed, both digitally following the μCT scan and physically both before and after enamel dissolution. The obtained measurements were subjected to statistical analysis through concordance coefficient measurements and linear regression. Results: A straight line correlation behavior with no statistically significant difference was found between both methods, with a concordance correlation coefficient of 97%. Conclusion: The digital, nondestructive, μCT-layered, three-dimensional reconstruction method presented comparable results to acid-etched enamel dissolution, confirming that both options are reliable for the histoanatomical analysis of enamel and dentin morphologies.