Becker Ann-Kristin, Erfle Holger, Gunkel Manuel, Beil Nina, Kaderali Lars, Starkuviene Vytaute
Institute of Bioinformatics, University Medicine Greifswald, 17475 Greifswald, Germany.
BioQuant, Heidelberg University, 69120 Heidelberg, Germany.
High Throughput. 2018 May 15;7(2):13. doi: 10.3390/ht7020013.
Multi-well plates and cell arrays enable microscopy-based screening assays in which many samples can be analysed in parallel. Each of the formats possesses its own strengths and weaknesses, but reference comparisons between these platforms and their application rationale is lacking. We aim to fill this gap by comparing two RNA interference (RNAi)-mediated fluorescence microscopy-based assays, namely epidermal growth factor (EGF) internalization and cell cycle progression, on both platforms. Quantitative analysis revealed that both platforms enabled the generation of data with the appearance of the expected phenotypes significantly distinct from the negative controls. The measurements of cell cycle progression were less variable in multi-well plates. The result can largely be attributed to higher cell numbers resulting in less data variability when dealing with the assay generating phenotypic cell subpopulations. The EGF internalization assay with a uniform phenotype over nearly the whole cell population performed better on cell arrays than in multi-well plates. The result was achieved by scoring five times less cells on cell arrays than in multi-well plates, indicating the efficiency of the cell array format. Our data indicate that the choice of the screening platform primarily depends on the type of the cellular assay to achieve a maximum data quality and screen efficiency.
多孔板和细胞阵列可实现基于显微镜的筛选分析，能够同时对多个样本进行分析。每种形式都有其自身的优缺点，但这些平台之间的参考比较及其应用原理尚付阙如。我们旨在通过在这两种平台上比较两种基于RNA干扰（RNAi）介导的荧光显微镜分析方法，即表皮生长因子（EGF）内化和细胞周期进程，来填补这一空白。定量分析表明，两种平台都能够生成数据，且预期表型的出现与阴性对照有显著差异。在多孔板中，细胞周期进程的测量结果变异性较小。这一结果很大程度上归因于细胞数量较多，在处理产生表型细胞亚群的分析时，数据变异性较小。在细胞阵列上，几乎整个细胞群体具有统一表型的EGF内化分析比在多孔板中表现更好。在细胞阵列上评分的细胞数量比在多孔板中少五倍，从而获得了这一结果，表明了细胞阵列形式的效率。我们的数据表明，筛选平台的选择主要取决于细胞分析的类型，以实现最大的数据质量和筛选效率。
