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Multispectral imaging and automated laser capture microdissection of human cortical neurons: A quantitative study of CXCR4 expression
Journal article   Open access

Multispectral imaging and automated laser capture microdissection of human cortical neurons: A quantitative study of CXCR4 expression

Jonathan Pitcher, Roberto Wurth, Saori Shimizu and Olimpia Meucci
Methods in molecular biology (Clifton, N.J.), v 1013, pp 31-48
2013
DOI: https://doi.org/10.1007/978-1-62703-426-5_3
PMID: 23625491
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https://doi.org/10.1007/978-1-62703-426-5_3View
Published, Version of Record (VoR) Open

Abstract

multispectral imaging chemokine quantitative LCM neuron CXCR4 human cortex immunohistochemistry laser capture microdissection
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 is commonly compounded by variations in post-mortem interval, formalin fixation time, and tissue processing among others. The result is a sample population that is inherently heterogeneous, requiring a single protocol that is sensitive to low levels of antigen while minimizing background and non-specific staining. Here, we describe a single immunohistochemistry (IHC) 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 (MSI) or 2) isolate neuronal RNA through automated laser capture microdissection (LCM).

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