使用PALM系统进行基因表达。

Gene expression using the PALM system.

作者信息

Lu Jian-Xin, Szeto Cheuk-Chun

机构信息

Department of Medicine & Therapeutics, Prince of Wales Hospital, The Chinese University of Hong Kong, Shatin, Hong Kong, China.

出版信息

Methods Mol Biol. 2011;755:47-56. doi: 10.1007/978-1-61779-163-5_3.

DOI:10.1007/978-1-61779-163-5_3

PMID:21761292

Abstract

The PALM Robot MicroBeam laser microdissection system can isolate specified cells from complex tissues section, in a rapid and precise manner. Combined with other methods, PALM may be used for gene expression elucidating the role of specialized cell type in physiological and pathological activity. This chapter describes the application of the PALM MicroBeam system to isolate RNA from cells in a complex tissue for subsequent gene expression analysis. Protocols show the steps from preparation of tissue samples to the final quantitative results. The process is articulated in several steps, each of which requires optimal choices in order to obtain reliable data from a limited number of cells (500-10,000 cells). Furthermore, the notes regarding tissue preparation, microdissection of the interested cells, are also emphasized.

摘要

PALM机器人微束激光显微切割系统能够以快速、精确的方式从复杂组织切片中分离出特定细胞。与其他方法相结合，PALM可用于阐明特定细胞类型在生理和病理活动中的作用的基因表达研究。本章介绍了PALM微束系统在从复杂组织中的细胞分离RNA以进行后续基因表达分析方面的应用。实验方案展示了从组织样本制备到最终定量结果的步骤。该过程分为几个步骤，每个步骤都需要做出最佳选择，以便从有限数量的细胞（500 - 10,000个细胞）中获得可靠的数据。此外，还强调了有关组织制备、感兴趣细胞的显微切割的注意事项。