Field Lori A, Deyarmin Brenda, Shriver Craig D, Ellsworth Darrell L, Ellsworth Rachel E
Windber Research Institute, Windber, PA, USA.
Methods Mol Biol. 2011;755:17-45. doi: 10.1007/978-1-61779-163-5_2.
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 micro-array data from high-quality frozen breast tissue specimens, as well as challenges associated with formalin-fixed paraffin-embedded specimens.
基于微阵列的基因表达谱分析正在彻底改变生物医学研究，它使人们能够在单个实验中对数千个基因的表达谱进行检测。在癌症研究中，使用激光显微切割（LM）从组织中分离RNA，能够准确识别构成肿瘤及其周围微环境的不同细胞类型的分子谱。由于RNA是一种不稳定的分子，从组织中提取的RNA质量会受到样品制备和处理的影响。因此，已经开发出特殊的方案来在激光显微切割后分离出研究级别的RNA。本章详细描述了用于从高质量冷冻乳腺组织标本生成微阵列数据的方案，以及与福尔马林固定石蜡包埋标本相关的挑战。
