Department of Chemical Engineering, Northeastern University, Boston, MA, 02115, USA.
Department of Chemical Engineering, Northeastern University, Boston, MA, 02115, USA.
Anal Chim Acta. 2019 Aug 13;1065:56-63. doi: 10.1016/j.aca.2019.02.040. Epub 2019 Mar 2.
Toxicity assay is crucial in identifying biological mechanisms, detecting disease and screening therapeutics. Single cell toxicity assay reveals heterogeneous responses of each cell without ensemble average, but still cannot indicate responses of the same individual cells to multiple treatments. This article reports an identical cell assay that can be used at high throughput to locate each cell and track its responses to multiple treatments (chemicals and radiation), thus greatly facilitates toxicity based drug screening and testing. Each cell is located through its coordinators pre-engraved on a patterned substrate with soft lithography. Cell responses to X-ray radiation, chemical reagents or a combination of both are obtained by probing reactive oxygen species (ROS) signals and quantified by fluorescent intensity in MATLAB. The method can provide toxicities of over thousands of identical cells with superior statistics power to allow deep analysis of toxicity data.
毒性分析在确定生物学机制、检测疾病和筛选治疗方法方面至关重要。单细胞毒性分析揭示了每个细胞的异质反应，而没有总体平均值，但仍然不能表明同一单个细胞对多种处理的反应。本文报道了一种可用于高通量的相同细胞分析方法，该方法可以定位每个细胞并跟踪其对多种处理（化学物质和辐射）的反应，从而极大地促进了基于毒性的药物筛选和测试。通过软光刻在具有图案的基底上预先刻划每个细胞的坐标来定位细胞。通过探测活性氧（ROS）信号获得细胞对 X 射线辐射、化学试剂或两者的组合的反应，并在 MATLAB 中通过荧光强度进行量化。该方法可以提供数千个相同细胞的毒性数据，具有较高的统计学功效，从而可以深入分析毒性数据。
