Laser Microdissection for Gene Expression Profiling
Microarray-based gene expression profiling is revolutionizing biomedical research by allowing expression profiles of thousands of genes to be interrogated in a single experiment. In cancer research, the use of laser microdissection (LM) to isolate RNA from tissues provides the ability to accurately identify molecular profiles from different cell types that comprise the tumor and its surrounding microenvironment. Because RNA is an unstable molecule, the quality of RNA extracted from tissues can be affected by sample preparation and processing. Thus, special protocols have been developed to isolate research-quality RNA after LM. This chapter provides detailed descriptions of protocols used to generate microarray data from high-quality frozen breast tissue specimens, as well as challenges associated with formalin-fixed paraffin-embedded specimens.
Key wordsLaser microdissection Gene expression Microarray Frozen tissue FFPE Molecular signature Breast cancer
This work was supported by the United States Department of Defense (Military Molecular Medicine Initiative MDA W81XWH-05-2-0075, protocol #01-20006) and was performed under the auspices of the Clinical Breast Care Project, a joint effort of many investigators and staff members. The opinion and assertions contained herein are the private views of the authors and are not to be construed as official or as representing the views of the Department of the Army or the Department of Defense.
- 1.Collins, F. S. and Barker, A. D. (2007) Mapping the cancer genome. Pinpointing the genes involved in cancer will help chart a new course across the complex landscape of human malignancies. Sci. Am. 296, 50–57.Google Scholar