High thickness histological sections as alternative to study the three-dimensional microscopic human sub-cortical neuroanatomy
Stereotaxy is based on the precise image-guided spatial localization of targets within the human brain. Even with the recent advances in MRI technology, histological examination renders different (and complementary) information of the nervous tissue. Although several maps have been selected as a basis for correlating imaging results with the anatomical locations of sub-cortical structures, technical limitations interfere in a point-to-point correlation between imaging and anatomy due to the lack of precise correction for post-mortem tissue deformations caused by tissue fixation and processing. We present an alternative method to parcellate human brain cytoarchitectural regions, minimizing deformations caused by post-mortem and tissue-processing artifacts and enhancing segmentation by means of modified high thickness histological techniques and registration with MRI of the same specimen and into MNI space (ICBM152). A three-dimensional (3D) histological atlas of the human thalamus, basal ganglia, and basal forebrain cholinergic system is displayed. Structure’s segmentations were performed in high-resolution dark-field and light-field microscopy. Bidimensional non-linear registration of the histological slices was followed by 3D registration with in situ MRI of the same subject. Manual and automated registration procedures were adopted and compared. To evaluate the quality of the registration procedures, Dice similarity coefficient and normalized weighted spectral distance were calculated and the results indicate good overlap between registered volumes and a small shape difference between them in both manual and automated registration methods. High thickness high-resolution histological slices in combination with registration to in situ MRI of the same subject provide an effective alternative method to study nuclear boundaries in the human brain, enhancing segmentation and demanding less resources and time for tissue processing than traditional methods.
KeywordsCytoarchitecture Thalamus Sub-cortical atlas Magnetic resonance imaging
The authors would like to thank the team participating on the São Paulo-Würzburg collaborative project. This includes all members of the Brain Bank of the Brazilian Aging Brain Research Group (BBBABSG) of the University of São Paulo Medical School, Mrs. E. Broschk and Mrs. A. Bahrke from the Morphological Brain Research Unit of the University of Würzburg, Germany.
Compliance with ethical standards
This study was supported by resources from the University of Sao Paulo School of Medicine, Brazil and University of Würzburg, Germany. The author Eduardo Joaquim Lopes Alho was supported by a scholarship from CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) agency, Brazil, for doctoral studies at the University of Würzburg, Germany. The authors do not have personal financial or institutional interest in any of the drugs, materials, or devices described in this article.
Conflict of interest
The authors disclose any actual or potential conflict of interest including any financial, personal, or other relationships with other people or organizations within 3 years of beginning the submitted work that could inappropriately influence, or be perceived to influence, their work.
The work described has been carried out in accordance with The Code of Ethics of the World Medical Association (Declaration of Helsinki).
Supplementary material 1 (WMV 26324 kb)
Supplementary material 2 (WMV 21876 kb)
Supplementary material 3 (WMV 188761 kb)
- Afshar F, Watkins ES, Yap JC (1978) Stereotaxic atlas of the human brainstem and cerebellar nuclei: a variability study. Raven Press, New YorkGoogle Scholar
- Alegro M, Loring B, Alho E et al (2016) Multimodal whole brain registration: MRI and high resolution histology. http://www.cv-foundation.org/openaccess/content_cvpr_2016_workshops/w15/papers/Alegro_Multimodal_Whole_Brain_CVPR_2016_paper.pdf. Accessed 9 Sep 2016
- Alic L, Haeck JH (2010) Multi-modal image registration: matching MRI with histology. SPIE Medical Imaging, San DiegoGoogle Scholar
- Andrew J, Watkins ES (1969) A stereotaxic atlas of the human thalamus and adjacent structures: a variability study. Williams and Wilkins, BaltimoreGoogle Scholar
- Bauchot R (1967) Modifications of brain weight in the course of fixation. J Für Hirnforsch 9:253–283Google Scholar
- Emmers R, Tasker R (1975) The human somesthetic thalamus: with maps for physiological target localization during stereotactic neurosurgery. Raven Press, New YorkGoogle Scholar
- Franklin K, Paxinos G (2008) The mouse brain in stereotaxic coordinates. Academic Press, New YorkGoogle Scholar
- Hassler R, Schaltenbrand G, Walker E (1982) Architectonic organization of the thalamic nuclei. In: Schaltenbrand G, Walker E (eds) Stereotaxy of the human brain: anatomical, physiological and clinical applications, 2nd edn. George Thieme Verlag, StuttgartGoogle Scholar
- Heinsen H, Arzberger T, Schmitz C (2000) Celloidin mounting (embedding without infiltration)—a new, simple and reliable method for producing serial sections of high thickness through complete human brains and its application to stereological and immunohistochemical investigations. J Chem Neuroanat 20:49–59CrossRefPubMedGoogle Scholar
- König JFR, Klippel RA (1963) The rat brain: a stereotaxic atlas of the forebrain and lower parts of the brain stem. Williams and Wilkins, BaltimoreGoogle Scholar
- Kumazawa-Manita N, Katayama M, Hashikawa T, Iriki A (2013) Three-dimensional reconstruction of brain structures of the rodent Octodon degus: a brain atlas constructed by combining histological and magnetic resonance images. Exp Brain Res 231:65–74. https://doi.org/10.1007/s00221-013-3667-1 CrossRefPubMedPubMedCentralGoogle Scholar
- Mai JK, Paxinos G, Voss T (2008) Atlas of the human brain. Academic Press, New YorkGoogle Scholar
- Nowinski WL, Fang A, Nguyen BT et al (1997) Multiple brain atlas database and atlas-based neuroimaging system. Comput Aided Surg Off J Int Soc Comput Aided Surg 2:42–66. https://doi.org/10.1002/(SICI)1097-0150(1997)2:1<42:AID-IGS7>3.0.CO;2-N CrossRefGoogle Scholar
- Nowinski WL, Belov D, Thirunavuukarasuu A, Benabid AL (2005) A probabilistic functional atlas of the VIM nucleus constructed from pre-, intra- and postoperative electrophysiological and neuroimaging data acquired during the surgical treatment of Parkinson’s disease patients. Stereotact Funct Neurosurg 83:190–196. https://doi.org/10.1159/000091082 CrossRefPubMedGoogle Scholar
- Ono M, Kubik S, Abernathey CD (1990) Atlas of the cerebral sulci. G. Thieme Verlag/Thieme Medical Publishers, Stuttgart/New YorkGoogle Scholar
- Paxinos G, Watson C (1986) The rat brain in stereotaxic coordinates. Academic Press, New YorkGoogle Scholar
- Saleem KS, Logothetis NK (2012) A combined MRI and histology atlas of the rhesus monkey brain in stereotaxic coordinates. Academic Press, New YorkGoogle Scholar
- Schaltenbrand G, Bailey P (1959) Introduction to stereotaxis with an atlas of the human brain, vol I: Text, vol II: Plate 1–57, vol III: Plate 58–76. Georg Thieme, Grune & Stratton:Stuttgart, New YorkGoogle Scholar
- Schaltenbrand G, Hassler R, Wahren W (1977) Atlas for stereotaxy of the human brain: with an accompanying guide. Thieme, StuttgartGoogle Scholar
- Schwarz AJ, Danckaert A, Reese T et al (2006) A stereotaxic MRI template set for the rat brain with tissue class distribution maps and co-registered anatomical atlas: application to pharmacological MRI. NeuroImage 32:538–550. https://doi.org/10.1016/j.neuroimage.2006.04.214 CrossRefPubMedGoogle Scholar
- Shanta TR, Manocha SL, Bourne GH (1968) A stereotaxic atlas of the java monkey brain (Macaca irus). Basel, Karger, doi: 10.1159/000390681
- Talairach J (1957) Atlas d’anatomie stéréotaxique; repérage radiologique indirect des noyaux gris centraux des régions mésencéphalo-sous-optique et hypothalamique de l’homme. Masson, ParisGoogle Scholar
- Talairach J, Tournoux P (1988) Co-planar stereotaxic atlas of the human brain: 3-dimensional proportional system: an approach to medical cerebral imaging. Thieme, StuttgartGoogle Scholar