Evensen Lasse, Link Wolfgang, Lorens James B
Department of Biomedicine, University of Bergen, Bergen, Norway.
Methods Mol Biol. 2013;931:139-51. doi: 10.1007/978-1-62703-056-4_8.
Automated multicolor fluorescence microscopy facilitates high-throughput quantitation of cellular parameters of complex, organotypic systems. In vitro co-cultured vascular cells form capillary-like networks that model facets of angiogenesis, making it an attractive alternative for anti-angiogenic drug discovery. We have adapted this angiogenesis assay system to a high-throughput format to enable automated image-based high-throughput screening of live primary human vascular cell co-cultures with chemical libraries for anti-angiogenic drug discovery. Protocols are described for setup of a fluorescence-based co-culture assay, live cell image acquisition, image analysis of morphological parameters, and screening data handling.
自动化多色荧光显微镜有助于对复杂的器官型系统的细胞参数进行高通量定量分析。体外共培养的血管细胞形成类似毛细血管的网络，模拟血管生成的各个方面，这使其成为抗血管生成药物发现的一个有吸引力的替代方案。我们已将这种血管生成检测系统调整为高通量形式，以便能够对活的原代人血管细胞共培养物与化学文库进行基于图像的自动化高通量筛选，用于抗血管生成药物发现。本文描述了基于荧光的共培养检测设置、活细胞图像采集、形态学参数的图像分析以及筛选数据处理的方案。
