Galbraith David W, Elumalai Rangasamy, Gong Fang Cheng
Department of Plant Sciences, University of Arizona, Tucson, AZ, USA.
Methods Mol Biol. 2004;263:259-80. doi: 10.1385/1-59259-773-4:259.
Flow cytometry and cell sorting provides unparalleled means for the identification and purification of specific cell types. It is a mature technology having been in existence commercially for the last 25 yr. High-throughput transcriptional profiling methods have emerged relatively recently. These provide the means to characterize efficiently the genome-wide contribution of individual genes to gene expression. A combination of these methods offers the opportunity to explore the relationship between gene expression and the ways in which different cell types are formed and maintained. This chapter provides a review of published methods for analysis of global gene expression within different cell types in complex tissues and organs, and provides practical details concerning microarray fabrication and use based on presynthesized 70-mer oligonucleotide array elements.
流式细胞术和细胞分选技术为特定细胞类型的识别与纯化提供了无与伦比的手段。它是一项成熟技术,在过去25年里一直有商业应用。高通量转录谱分析方法相对较新才出现。这些方法提供了有效表征单个基因对全基因组基因表达贡献的手段。将这些方法结合起来,有机会探索基因表达与不同细胞类型形成和维持方式之间的关系。本章综述了已发表的用于分析复杂组织和器官中不同细胞类型内全局基因表达的方法,并提供了基于预合成70聚体寡核苷酸阵列元件的微阵列制备和使用的实际细节。