title: A robust RNA integrity-preserving staining protocol for laser capture microdissection of endometrial cancer tissue.
abstract: Laser capture microdissection of frozen tissue sections allows homogeneous cell populations to be isolated for expression profiling. However, this requires striking a balance between retaining adequate morphology for accurate microdissection and maintaining RNA integrity. Various staining protocols were applied to frozen endometrial carcinoma tissue sections. Although alcohol-based methods were superior to aqueous stains for maintaining RNA integrity, they suffered from irreproducible staining intensity. We developed a modified alcohol-based, buffered cresyl violet staining protocol that provides reproducible staining with minimal RNA degradation suitable for tissues with moderate to high levels of intrinsic RNase activity.
authors.affiliation: Gynaeimmunology and Oncology Group, YCR and Liz Dawn Pathology and Translational Sciences Centre, Leeds Institute of Molecular Medicine, St. James's University Hospital, Leeds LS9 7TF, UK. medmic@leeds.ac.uk
authors.forename: Michele
authors.initials: M
authors.lastname: Cummings
authors.forename: Ciara V
authors.initials: CV
authors.lastname: McGinley
authors.forename: Nafisa
authors.initials: N
authors.lastname: Wilkinson
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Field
authors.forename: Sean R
authors.initials: SR
authors.lastname: Duffy
authors.forename: Nicolas M
authors.initials: NM
authors.lastname: Orsi
mesh_terms: D018269:Carcinoma, Endometrioid / Q000235:genetics / Q000473:pathology*; D016889:Endometrial Neoplasms / Q000235:genetics / Q000473:pathology*; D005260:Female; D005629:Frozen Sections; D006801:Humans; D007834:Lasers; D042282:Microdissection; D012334:RNA, Neoplasm / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D012334:RNA, Neoplasm