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Immunohistochemical staining of post-mortem adult human brain sections

Nature Protocols volume 1, pages 2719–2732 (2006)Cite this article

Abstract

One of the challenges for modern neuroscience is to understand the basis of coordinated neuronal function and networking in the human brain. Some of these questions can be addressed using low- and high-resolution imaging techniques on post-mortem human brain tissue. We have established a versatile protocol for fixation of post-mortem adult human brain tissue, storage of the tissue in a human brain bank, and immunohistochemical analysis in order to understand human brain functions in normal controls and in neuropathological conditions. The brains are fixed by perfusion through the internal carotid and basilar arteries to enhance the penetration of fixative throughout the brain, then blocked, postfixed, cryoprotected, snap-frozen and stored at –80 °C. Sections are processed for immunohistochemical single- or double-label staining and conventional-, electron- or confocal laser scanning-microscopy analysis. The results gained using this tissue and protocol are vital for determining the localization of neurochemicals throughout the human brain and to document the changes that occur in neurological diseases.

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Figure 1: Examples of images taken of sections from various regions of the human brain that have been immunohistochemically labeled with a variety of neurotransmitter receptors and neurochemical labels.
Figure 2: Confocal laser-scanning microscopy images of the human subventricular zone (SVZ) showing progenitor cells that can be detected using specific antibodies.
Figure 3: Illustrations showing the procedure for blocking of the brain.
Figure 4: Blocking the basal ganglia, thalamus and hippocampus.
Figure 5

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Acknowledgements

We would like to thank J. Bullock for expert technical assistance, the Biomedical Imaging Research Unit of the University of Auckland for their expert assistance and all of the families for donating the tissue for our studies. This study was supported by the Health Research Council of New Zealand, The Neurological Foundation of New Zealand and the Auckland Medical Research Fund. M.C is supported by a Neurological Foundation of New Zealand Wrightson Fellowship.

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Authors and Affiliations

  1. Department of Anatomy with Radiology, Faculty of Medical and Health Science, University of Auckland, Private Bag, 92019, Auckland, New Zealand

    Henry J Waldvogel, Maurice A Curtis & Richard L M Faull

  2. Institute of Neurosciences and Physiology at Sahlgrenska Academy, Göteborg University, Medicinaregat 11, Box 432, Göteborg, s-405 30, Sweden

    Maurice A Curtis

  3. Molecular Neuroscience, School of Medicine, University of Wales Swansea, Singleton Park, Swansea, SA2 8PP, Wales, U.K.

    Kristin Baer & Mark I Rees

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  1. Henry J Waldvogel
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Correspondence to Henry J Waldvogel.

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Waldvogel, H., Curtis, M., Baer, K. et al. Immunohistochemical staining of post-mortem adult human brain sections. Nat Protoc 1, 2719–2732 (2006). https://doi.org/10.1038/nprot.2006.354

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