Immunohistochemical Detection of Alpha-Synuclein in Unfixed Human Brain Tissue
In both research and diagnostics, immunohistochemistry is an essential method for assessing pathology in neurodegenerative diseases. Typically, at autopsy, one hemisphere of the brain is formalin fixed for sectioning and histochemical analysis, while the other hemisphere is flash frozen for biochemical studies. While this process ensures the generation of high-quality slides, it is important to assess the pathology of tissue near pieces used for biochemical analyses, since pathology burden can be unevenly distributed throughout the brain (Fig. 1). Here, we describe a method for assessing alpha-synuclein (αSyn) pathology in unfixed human brain tissue using a novel monoclonal antibody.
Key wordsAlpha-synuclein Immunohistochemistry Human tissue Frozen Fresh
The author would like to thank Dr. Mel Feany, Dr. Liqun Wang, and Kit Sin Tuen from the Feany Laboratory at the Ann Romney Center for Neurologic Disease at Brigham and Women’s Hospital for assistance with developing the protocol. The author would also like to acknowledge the Pathology Core at the Dana-Farber Cancer Institute for their work in optimizing the 2F12 antibody concentration.
- 9.Childs, G (2014) History of immunohistochemistry. In: McManus LM, Mitchell RN (ed) Pathobiology of human disease, 1. Academic Press, Cambridge, MAGoogle Scholar