Abstract
Quantifying protein and RNA expression within specific cell populations in vivo is an essential step in unraveling the complex mechanisms of neurological disease. The challenges associated with studying human brain tissue are commonly compounded by variations in postmortem interval, formalin fixation time, and tissue processing methods among others. The result is a sample population that is inherently heterogeneous, implying the need for reliable protocols that are sensitive to low levels of antigen while minimizing background and nonspecific staining. Here, we describe a single immunohistochemistry protocol on formalin-fixed, paraffin-embedded human cortex which can be adapted to (1) quantify the relative protein expression of the chemokine receptor, CXCR4, using multispectral image or (2) isolate neuronal RNA through automated laser capture microdissection.
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References
Cattoretti G, Pileri S, Paravicini C, Becker MH, Poggi S, Bifulco C, Key G, D’Amato L, Sabattin E, Feudale E (1993) Antigen unmasking on formalin-fixed, paraffin-embedded tissue sections. J Pathol 171:83–98
Boenisch T (2005) Effect of heat induced antigen retrieval following inconsistent formalin fixation. Appl Immunohistochem Mol Morphol 13:283–286
Buchwalow I, Bocker W (2010) Immuno-histochemistry: basics and methods. Springer-Verlag, Berlin/Heidelberg
Acknowledgements
This work was supported by National Institute of Health—National Institute on Drug Abuse Grants DA15014 and DA19808 (OM). Jonathan Pitcher was a student fellow of the neuroAIDS training grant Interdisciplinary and Translational Research Training Program in neuroAIDS (T32MH078795).
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Pitcher, J., Wurth, R., Shimizu, S., Meucci, O. (2013). Multispectral Imaging and Automated Laser Capture Microdissection of Human Cortical Neurons: A Quantitative Study of CXCR4 Expression. In: Cardona, A., Ubogu, E. (eds) Chemokines. Methods in Molecular Biology, vol 1013. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-426-5_3
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DOI: https://doi.org/10.1007/978-1-62703-426-5_3
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-425-8
Online ISBN: 978-1-62703-426-5
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