Stereological techniques using point counting and planimetry have been used to estimate pituitary gland volume. However, many studies have estimated pituitary gland volume by the mathematical approach the elliptic formula. The objective of the current study was to determine pituitary gland volume using stereological methods and elliptic formula on magnetic resonance imaging (MRI). In this study, pituitary gland volumes were estimated in a total of 28 subjects (22 females, 6 males,) who were free of any pituitary or neurological symptoms and signs. The mean ± SD pituitary gland volumes for the point counting, planimetry and elliptic formulae groups were 582.14 ± 140.16 mm³, 610.08 ± 133.17 mm³, and 432.82 ± 147.38 mm³, respectively. The mean CE for the pituitary gland volume estimates derived from the point counting technique was 8.07%. No significant difference was found between point counting and planimetric methods for the pituitary gland volume (P > 0.05). In addition, there was a 26.14 and 29.71% underestimation of pituitary volume as measured by the elliptic formula compared to the point counting and planimetric techniques, respectively. From these results, it can be concluded that stereological techniques are unbiased, efficient and reliable methods and are ideally suitable for in vivo examination of MRI data for pituitary gland volume estimation. Hence, we suggest that estimating pituitary gland volume using MRI study and stereology may be clinically relevant for pituitary surgeons for the investigation of the structure and function of the pituitary gland.