title: Laser-capture microdissection.
abstract: Deciphering the cellular and molecular interactions that drive disease within the tissue microenvironment holds promise for discovering drug targets of the future. In order to recapitulate the in vivo interactions thorough molecular analysis, one must be able to analyze specific cell populations within the context of their heterogeneous tissue microecology. Laser-capture microdissection (LCM) is a method to procure subpopulations of tissue cells under direct microscopic visualization. LCM technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity (LOH) analysis, RNA transcript profiling, cDNA library generation, proteomics discovery and signal-pathway profiling. Herein we provide a thorough description of LCM techniques, with an emphasis on tips and troubleshooting advice derived from LCM users. The total time required to carry out this protocol is typically 1-1.5 h.
authors.affiliation: Center for Applied Proteomics and Molecular Medicine, George Mason University, 10900 University Blvd. MS 4E3, Manassas, Virginia, USA.
authors.forename: Virginia
authors.initials: V
authors.lastname: Espina
authors.forename: Julia D
authors.initials: JD
authors.lastname: Wulfkuhle
authors.forename: Valerie S
authors.initials: VS
authors.lastname: Calvert
authors.forename: Amy
authors.initials: A
authors.lastname: VanMeter
authors.forename: Weidong
authors.initials: W
authors.lastname: Zhou
authors.forename: George
authors.initials: G
authors.lastname: Coukos
authors.forename: David H
authors.initials: DH
authors.lastname: Geho
authors.forename: Emanuel F
authors.initials: EF
authors.lastname: Petricoin
authors.forename: Lance A
authors.initials: LA
authors.lastname: Liotta
mesh_terms: D002469:Cell Separation / Q000295:instrumentation / Q000379:methods; D004247:DNA; D007259:Infrared Rays; D007834:Lasers; D042282:Microdissection / Q000295:instrumentation* / Q000379:methods*; D011506:Proteins; D012313:RNA; D013194:Staining and Labeling; D014466:Ultraviolet Rays
chemical_list: D011506:Proteins; D012313:RNA; D004247:DNA