Castro Nadia P, Merchant Anand S, Saylor Karen L, Anver Miriam R, Salomon David S, Golubeva Yelena G
Tumor Growth Factor Section, Mouse Cancer Genetics Program, National Cancer Institute, Frederick, MD, 21702, United States of America.
CCRIFX Bioinformatics Core, National Cancer Institute, Bethesda, MD, 20892, United States of America.
PLoS One. 2016 Apr 14;11(4):e0153270. doi: 10.1371/journal.pone.0153270. eCollection 2016.
Laser capture microdissection (LCM) of tissue is an established tool in medical research for collection of distinguished cell populations under direct microscopic visualization for molecular analysis. LCM samples have been successfully analyzed in a number of genomic and proteomic downstream molecular applications. However, LCM sample collection and preparation procedure has to be adapted to each downstream analysis platform. In this present manuscript we describe in detail the adaptation of LCM methodology for the collection and preparation of fresh frozen samples for NanoString analysis based on a study of a model of mouse mammary gland carcinoma and its lung metastasis. Our adaptation of LCM sample preparation and workflow to the requirements of the NanoString platform allowed acquiring samples with high RNA quality. The NanoString analysis of such samples provided sensitive detection of genes of interest and their associated molecular pathways. NanoString is a reliable gene expression analysis platform that can be effectively coupled with LCM.
组织的激光捕获显微切割（LCM）是医学研究中的一种成熟工具，用于在直接显微镜观察下收集特定细胞群体以进行分子分析。LCM样本已在许多基因组和蛋白质组下游分子应用中成功分析。然而，LCM样本收集和制备程序必须适应每个下游分析平台。在本手稿中，我们基于对小鼠乳腺癌及其肺转移模型的研究，详细描述了LCM方法在收集和制备用于NanoString分析的新鲜冷冻样本方面的适应性。我们将LCM样本制备和工作流程调整以适应NanoString平台的要求，从而能够获取具有高RNA质量的样本。对此类样本的NanoString分析能够灵敏地检测感兴趣的基因及其相关分子途径。NanoString是一个可靠的基因表达分析平台，可与LCM有效结合。
