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: Michele Cummings, Ciara V McGinley, Nafisa Wilkinson, Sarah L Field, Sean R Duffy, Nicolas M Orsi
• Journal: Analytical biochemistry (Anal Biochem) Vol. 416 Issue 1 Pg. 123-5 (Sep 01 2011) ISSN: 1096-0309 [Electronic] United States
• PMID: 21624343 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2011 Elsevier Inc. All rights reserved.

Chemical References

• RNA, Neoplasm

Topics

• Carcinoma, Endometrioid (genetics, pathology)
• Endometrial Neoplasms (genetics, pathology)
• Female
• Frozen Sections
• Humans
• Lasers
• Microdissection
• RNA, Neoplasm (analysis, genetics, metabolism)
• Staining and Labeling (methods)