Background As an advanced imaging technique for the human brain, the importance of magnetic resonance imaging technique (MRI) is indisputable. The study aims to contribute to the literature by imaging post-mortem human brain hemispheres fixed with the Klinger method through the a 3.0 Tesla MRI Scanner and by defining the supratentorial major white matter tracts and central core anatomical structures. Methods In our study, 10 post-mortem human brain hemisphere specimens were placed in 10% formalin solution for at least two months according to the Klingler method. The images were obtained using a 3.0 Tesla MRI Scanner. Anatomical structures were described on the T1-T2 axial, coronal, and sagittal MRI sections and compared with control images obtained from healthy humans. Results Our examination revealed major association fibers, the basal cores and nuclei were denser, and the connections between them were clearly visible. The basal nuclei particularly were visualized more clearly compared with the normal MRI examinations. The claustrum, putamen, lateral and medial part of globus pallidus, and the caudolenticular bridges of the caudate nucleus could be clearly distinguished. The optic radiation line toward the occipital area as well as the forceps major and minor were distinct in the axial sections. Meanwhile, the imaging emphasized the importance of temporal stem, and the fibers it contained were clearly observed in the coronal sections. Conclusion The use of hemispheres fixed using the Klinger method in post-mortem MRI examinations on brain hemispheres showed a clear separation of white matter fibers and nuclear structures.