La Spada Alberto, Rainoldi Barnaba, De Blasio Andrea, Biunno Ida
UOS-IRGB-CNR, Via Fantoli 16/15, 20138, Milano, Italy.
IRCCS-Multimedica, Via Fantoli 16/15, 20138, Milano, Italy.
Microarrays (Basel). 2014 Jun 26;3(3):159-67. doi: 10.3390/microarrays3030159.
There is virtually an unlimited number of possible Tissue Microarray (TMA) applications in basic and clinical research and ultimately in diagnostics. However, to assess the functional importance of novel markers, researchers very often turn to cell line model systems. The appropriate choice of a cell line is often a difficult task, but the use of cell microarray (CMA) technology enables a quick screening of several markers in cells of different origins, mimicking a genomic-scale analysis. In order to improve the morphological evaluations of the CMA slides we harvested the cells by conventional trypsinization, mechanical scraping and cells grown on coverslips. We show that mechanical scraping is a good evaluation method since keeps the real morphology very similar to those grown on coverslips. Immunofluorescence images are of higher quality, facilitating the reading of the biomarker cellular and subcellular localization. Here, we describe CMA technology in stem cell research.
在基础研究、临床研究以及最终的诊断中,组织微阵列(TMA)实际上有着无数可能的应用。然而,为了评估新型标志物的功能重要性,研究人员常常求助于细胞系模型系统。选择合适的细胞系往往是一项艰巨的任务,但细胞微阵列(CMA)技术的使用能够在不同来源的细胞中快速筛选多种标志物,模拟基因组规模的分析。为了改进CMA载玻片的形态学评估,我们通过传统的胰蛋白酶消化法、机械刮除法以及在盖玻片上生长的细胞来收获细胞。我们表明机械刮除法是一种很好的评估方法,因为它能使真实形态与在盖玻片上生长的细胞非常相似。免疫荧光图像质量更高,便于读取生物标志物在细胞和亚细胞水平的定位。在此,我们描述干细胞研究中的CMA技术。