In recent years interest in the anatomy of corpus callosum (CC) has led to a number of studies on morphometric analysis of CC, either in vivo on magnetic resonance imaging (MRI) or on preserved (formalin-fixed) cadaveric brain specimens. There are very few reports comparing the values obtained by both these methodologies, and to the authors’ knowledge no study of CC morphometry in fresh (unfixed) autopsy brains has been done. Morphometric measurements of the CC were done on fresh autopsy brains (n = 15), preserved cadaveric brains (n = 44) and MRI scans (n = 30) in adults (age range: 20-60 years) of both sexes without any intracranial brain pathology, in north-west Indian subjects. Most of the CC measurements were found to be more in the fresh autopsy brain group as compared to the preserved cadaveric brain and MRI group. The distance between the splenium and superior colliculus in the fresh autopsy brain group was almost twice that in the other two groups. In addition, there was a similarity between the preserved cadaveric brain and MRI values for most of the parameters, but the distances between the genu and fornix and between the genu and anterior commissure were larger in the MRI group as compared to the other two groups. The present data may be of value in studying deviations from the normal in various disease processes as well in surgical planning of intraventricular and callosal pathologies.