1 Karlsruhe Institute of Technology, Institute of Toxicology and Genetics, Eggenstein-Leopoldshafen, Germany.
2 Karlsruhe Institute of Technology, Institute for Applied Computer Science, Eggenstein-Leopoldshafen, Germany.
SLAS Technol. 2017 Apr;22(2):163-175. doi: 10.1177/2211068216677204. Epub 2016 Nov 15.
Phenotypic cell-based high-throughput screenings play a central role in drug discovery and toxicology. The main tendency in cell screenings is the increase of the throughput and decrease of reaction volume in order to accelerate the experiments, reduce the costs, and enable screenings of rare cells. Conventionally, cell-based assays are performed in microtiter plates, which exist in 96- to 1536-wells formats and cannot be further miniaturized. In addition, performing screenings of suspension cells is associated with risk of losing cell content during the staining procedures and incompatibility with high-content microscopy. Here, we evaluate the Droplet-Microarray screening platform for culturing, screening, and imaging of suspension cells. We demonstrate pipetting-free cell seeding and proliferation of cells in individual droplets of 3-80 nL in volume. We developed a methodology to perform parallel treatment, staining, and fixation of suspension cells in individual droplets. Automated imaging of live suspension cells directly in the droplets combined with algorithms for pattern recognition for image analysis is demonstrated. We evaluated the developed methodology by performing a dose-response study with antineoplastic drugs. We believe that the DMA screening platform carries great potential to be adopted for broad spectrum of screenings of suspension cells.
表型细胞高通量筛选在药物发现和毒理学中起着核心作用。细胞筛选的主要趋势是增加通量和减少反应体积,以加速实验、降低成本,并实现稀有细胞的筛选。传统上,基于细胞的测定是在微孔板中进行的,微孔板有 96 孔到 1536 孔的格式,不能进一步小型化。此外,悬浮细胞的筛选与在染色过程中丢失细胞内容物的风险以及与高内涵显微镜不兼容有关。在这里,我们评估了用于悬浮细胞培养、筛选和成像的液滴微阵列筛选平台。我们证明了可以在体积为 3-80nL 的单个液滴中进行无移液细胞播种和增殖。我们开发了一种在单个液滴中对悬浮细胞进行平行处理、染色和固定的方法。直接在液滴中对活悬浮细胞进行自动成像,并结合用于图像分析的模式识别算法进行了演示。我们通过进行抗肿瘤药物的剂量反应研究来评估所开发的方法。我们相信,DMA 筛选平台具有广泛应用于悬浮细胞筛选的巨大潜力。