title: Region-specific in situ hybridization-guided laser-capture microdissection on postmortem human brain tissue coupled with gene expression quantification.
abstract: This chapter describes the procedure of in situ hybridization-guided laser-capture microdissection performed on postmortem human brain tissue. This procedure permits the precise collection of brain tissue within anatomically defined brain nuclei that is enriched with mRNA. The chapter emphasizes the specific handling of postmortem tissue and preservation of RNA integrity to ensure high-quality gene profiling. Downstream procedures including mRNA amplification, gene profiling using high-density microarray chips, and confirmation with quantitative real-time polymerase chain reaction (qPCR) are described. PCR primer design and cDNA quantification required for qPCR are delineated.
authors.affiliation: Centrum für Anatomie, Institut für Integrative Neuroanatomie, Charité Campus Mitte-Universitätsmedizin Berlin, Berlin, Germany. rbbernard@gmail.com
authors.forename: René
authors.initials: R
authors.lastname: Bernard
authors.forename: Sharon
authors.initials: S
authors.lastname: Burke
authors.forename: Ilan A
authors.initials: IA
authors.lastname: Kerman
mesh_terms: D019921:Biotinylation / Q000379:methods; D001921:Brain / Q000473:pathology*; D002138:Calibration; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D008867:Microtomy / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000235:genetics / Q000302:isolation & purification
chemical_list: D012313:RNA