Mustafa Dana, Kros Johan M, Luider Theo
Department of Pathology, Josephine Nefkens Institute, Erasmus Medical Center, Rotterdam, The Netherlands.
Methods Mol Biol. 2008;428:159-78. doi: 10.1007/978-1-59745-117-8_9.
Laser microdissection is an effective technique to harvest pure cell populations from complex tissue sections. In addition to using the microdissected cells in several DNA and RNA studies, it has been shown that the small number of cells obtained by this technique can also be used for proteomics analysis. Combining laser capture microdissection and different types of mass spectrometers opened ways to find and identify proteins that are specific for various cell types, tissues, and their morbid alterations. Although the combination of microdissection followed by the currently available techniques of proteomics has not yet reached the stage of genome wide representation of all proteins present in a tissue, it is a feasible way to find significant differentially expressed proteins in target tissues. Recent developments in mass spectrometric detection followed by proper statistics and bioinformatics enable to analyze the proteome of not more than 100-200 cells. Obviously, validation of result is essential. The present review describes and discusses the various methods developed to target cell populations of interest by laser microdissection, followed by analysis of their proteome.
激光显微切割是从复杂组织切片中获取纯细胞群体的有效技术。除了在多项DNA和RNA研究中使用显微切割的细胞外,研究表明,通过该技术获得的少量细胞也可用于蛋白质组学分析。将激光捕获显微切割与不同类型的质谱仪相结合,为发现和鉴定各种细胞类型、组织及其病变改变所特有的蛋白质开辟了道路。尽管显微切割与当前可用的蛋白质组学技术相结合尚未达到在组织中全面呈现所有蛋白质的基因组水平,但它是在靶组织中寻找显著差异表达蛋白质的可行方法。质谱检测的最新进展,再加上适当的统计学和生物信息学方法,能够分析不超过100 - 200个细胞的蛋白质组。显然,结果验证至关重要。本综述描述并讨论了通过激光显微切割靶向感兴趣的细胞群体,随后分析其蛋白质组所开发的各种方法。