Ruetze Martin, Lochner Katrin, Gallinat Stefan, Knott Anja
Beiersdorf AG, R&D, Skin Research Center, Hamburg, Germany.
Methods Mol Biol. 2010;585:183-92. doi: 10.1007/978-1-60761-380-0_14.
Epithelial tissues exhibit optimal conditions for studying cellular differentiation since the differentiation status of a single cell can be determined by its distance to the basal membrane. For that reason Laser Capture Microdissection (LCM) may serve as a perfect tool to compare the characteristics of cells that have been collected from different strata of the epithelium. However, as cell boundaries are not visible in untreated tissue sections, samples have to be stained to allow for sufficient structural orientation. This usually results in a considerable reduction of RNA content in the dissected specimen. To circumvent this problem, we have established a modified hematoxylin/eosin staining protocol that concurrently allows visualization of important structures and the subsequent isolation of sufficient RNA amounts to be used for linear amplification and quantitative analyses.
上皮组织为研究细胞分化提供了理想条件，因为单个细胞的分化状态可由其与基底膜的距离来确定。因此，激光捕获显微切割技术（LCM）可能是比较从上皮组织不同层次收集的细胞特征的完美工具。然而，由于在未经处理的组织切片中细胞边界不可见，样本必须进行染色以获得足够的结构定位。这通常会导致所切割标本中的RNA含量大幅降低。为了解决这个问题，我们建立了一种改良的苏木精/伊红染色方案，该方案能够同时使重要结构可视化，并随后分离出足够量的RNA用于线性扩增和定量分析。
