title: Multispectral imaging and automated laser capture microdissection of human cortical neurons: a quantitative study of CXCR4 expression.
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.
authors.affiliation: Department of Pharmacology, Drexel University College of Medicine, Philadelphia, PA, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Pitcher
authors.forename: Roberto
authors.initials: R
authors.lastname: Wurth
authors.forename: Saori
authors.initials: S
authors.lastname: Shimizu
authors.forename: Olimpia
authors.initials: O
authors.lastname: Meucci
mesh_terms: D057205:Automation, Laboratory; D002540:Cerebral Cortex / Q000737:chemistry* / Q000166:cytology; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D061346:Laser Capture Microdissection; D008853:Microscopy; D009474:Neurons / Q000737:chemistry*; D016612:Paraffin Embedding; D012313:RNA / Q000302:isolation & purification; D019718:Receptors, CXCR4 / Q000032:analysis* / Q000235:genetics; D016707:Tissue Fixation
chemical_list: C527276:CXCR4 protein, human; D005404:Fixatives; D019718:Receptors, CXCR4; D005557:Formaldehyde; D012313:RNA