{"paper_id":"b5aefbf4-3886-4ebd-8909-515ad62d67a6","body_text":"Abstract\nEndometriosis, a common gynecological disorder responsible for infertility and pelvic pain, is defined as the presence of endometrial glands and stroma within extra-uterine sites. Gene expression studies performed on endometriotic tissue homogenates have yielded results reflecting mRNA abundance in a mixture of cell types (including epithelial cells, stromal cells, fibrotic tissue, and muscle tissue). Therefore, a method for quantifying gene expression separately in individual cell populations is essential for identifying genetic markers. Laser capture microdissection is a technique for obtaining pure populations of cells from heterogeneous tissues. This chapter provides methods to obtain high-quality RNA suitable for a variety of different down stream applications from frozen endometrial and endometriotic tissues for laser capture microdissection, using the Arcturus PixCell II system.\nAccess this chapter\nTax calculation will be finalised at checkout\nPurchases are for personal use only\nSimilar content being viewed by others\nReferences\nMatsuzaki S, Canis M, Vaurs-Barriere C, Pouly JL, Boespflug-Tanguy O, Penault-Llorca F, Dechelotte P, Dastugue B, Okamura K, Mage G. (2004) DNA microarray analysis of gene expression profiles in deep endometriosis using laser capture microdissection. Mol Hum Reprod. 10:719–28.\nMatsuzaki S, Canis M, Vaurs-Barriere C, Boespflug-Tanguy O, Dastugue B, Mage G. (2005) DNA microarray analysis of gene expression in eutopic endometrium from patients with deep endometriosis using laser capture microdissection. Fertil Steril, 84 Suppl 2:1180–90.\nEmmert-Buck, M.R., Bonner, R.F., Smith, P.D., Chuaqui, R.F., Zhuang, Z., Goldstein, S.R., Weiss, R.A. and Liotta, L.A. (1996) Laser capture microdissection. Science, 274, 998–1001.\nFleige S, Pfaffl MW. (2006) RNA integrity and the effect on the real-time qRT-PCR performance. Mol Aspects Med. 2006; 27:126–39.\nFleige S, Walf V, Huch S, Prgomet C, Sehm J, Pfaffl MW (2006). Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR. Biotechnol Lett. 28:1601–13.\nSchroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, Lightfoot S, Menzel W, Granzow M, Ragg T (2006). The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol. 7:3.\nStrand C, Enell J, Hedenfalk I, Fernö M (2007). RNA quality in frozen breast cancer samples and the influence on gene expression analysis––a comparison of three evaluation methods using microcapillary electrophoresis traces. BMC Mol Biol. 8:38.\nGassmann, M. (2003) Quality Assurance of RNA derived from laser microdissected tissue samples obtained by the PALM(R) MicroBeam System using the RNA 6000 Pico LabChip(R) kit. 1-8. Agilent Technologies.\nAcknowledgments\nThis work was supported in part by Karl Storz Endoscopy & GmbH (Tuttlingen, Germany) and le Conseil Régional d’Auvergne (Recherche et Innovation Technologique) (Clermont-Ferrand, France).\nAuthor information\nAuthors and Affiliations\nEditor information\nEditors and Affiliations\nRights and permissions\nCopyright information\n© 2009 Humana Press, a part of Springer Science+Business Media, LLC\nAbout this protocol\nCite this protocol\nMatsuzaki, S., Canis, M., Mage, G. (2009). Use of Laser Capture Microdissection in Studying Hormone-Dependent Diseases: Endometriosis. In: Park-Sarge, OK., Curry, T. (eds) Molecular Endocrinology. Methods in Molecular Biology, vol 590. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-378-7_19\nDownload citation\nDOI: https://doi.org/10.1007/978-1-60327-378-7_19\nPublished:\nPublisher Name: Humana Press, Totowa, NJ\nPrint ISBN: 978-1-60327-377-0\nOnline ISBN: 978-1-60327-378-7\neBook Packages: Springer Protocols","source_license":"public-domain-us","license_restricted":false}