The hippocampal formation possesses an important role in the development and maintenance of short-term memory. In this study, magnetic resonance imaging (MRI) and gross histology were used to quantify the volume of the hippocampal formation in canines. High resolution MRI, using 1 mm thick slices and an intraplanar resolution of 0.35 mm was performed at 2.0 T both in vivo and in vitro following in situ fixation. The volumes of the hippocampal formations were determined from MR images and compared to those obtained from one mm thick gross histologic sections. The average volume of the canine hippocampal formation, measured from in vivo and in vitro MR images was 476.0 +/- 79.5 and 467.3 +/- 53.7 mm3, respectively. Determined from gross histology, the volume of the hippocampal formation was 463.6 +/- 24.1 mm3. Quantitation of the canine hippocampal formation using in vivo MRI showed good correlation with in vitro MRI and histology, verifying the reliability and reproducibility of in vivo MRI measurements. High resolution MRI using 1 mm thick slices through the whole canine hippocampal formation is necessary for accurate volume determination of a structure of this size.