title: Laser capture microdissection applications in breast cancer proteomics.
abstract: Breast cancer tissues are characterized by cellular heterogeneity, representing a mixture of, e.g., healthy epithelial ducts, invasive or in situ tumor cells, surrounding stroma, infiltrating immune cells, blood vessels, and capillaries. As a consequence, protein extracts from whole tissue lysates also represent a variety of cell types present in the tissues under examination. This, however, seriously hampers the analysis of tumor cell-specific signals, which is of interest when performing biomarker discovery-type of studies. Therefore, laser capture microdissection is a perfect tool to isolate a relatively pure population of cells of interest, such as tumor cells. In this chapter, we describe the use of the PALM MicroBeam system for laser microdissection and pressure catapulting. Protocols are provided for sectioning, staining, microdissection, sample preparation, and mass spectrometric analysis of snap frozen breast cancer tissue.
authors.affiliation: Department of Medical Oncology, Center for Translational Molecular Medicine, and Cancer Genomics Centre, Erasmus MC Rotterdam, Rotterdam, The Netherlands.
authors.forename: René B H
authors.initials: RB
authors.lastname: Braakman
authors.forename: Theo M
authors.initials: TM
authors.lastname: Luider
authors.forename: John W M
authors.initials: JW
authors.lastname: Martens
authors.forename: John A
authors.initials: JA
authors.lastname: Foekens
authors.forename: Arzu
authors.initials: A
authors.lastname: Umar
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D002853:Chromatography, Liquid / Q000379:methods; D005260:Female; D006801:Humans; D007834:Lasers; D042282:Microdissection / Q000295:instrumentation / Q000379:methods*; D008867:Microtomy / Q000379:methods; D010446:Peptide Fragments / Q000737:chemistry; D020543:Proteome / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics; D012984:Software; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013194:Staining and Labeling / Q000379:methods
chemical_list: D010446:Peptide Fragments; D020543:Proteome