We evaluated the accuracy of estimating the volume of biological soft tissues from their three-dimensional (3D) computer wireframe models, reconstructed from histological data sets obtained from guinea-pig spinal cords. We compared quantification from two methods of three-dimensional surface reconstruction to standard quantitative techniques, Cavalieri method employing planimetry and point counting and Geometric Best-Fitting. This involved measuring a group of spinal cord segments and test objects to evaluate the accuracy of our novel quantification approaches. Once a quantitative methodology was standardized there was no statistical difference in volume measurement of spinal segments between quantification methods. We found that our 3D surface reconstructions’ ability to model precisely actual soft tissues provided an accurate volume quantification of complex anatomical structures as standard approaches of Cavalieri estimation and Geometric Best-Fitting. Additionally, 3D reconstruction quantitatively interrogates and three-dimensionally images spinal cord segments and obscured internal pathological features with approximately the same effort required for standard quantification alone.