Department of Chemical Engineering, Northeastern University, Boston, MA 02115, USA.
Department of Chemical and Biomedical Engineering, Florida State University, Tallahassee, FL 32310, USA.
Anal Chim Acta. 2018 May 12;1007:26-32. doi: 10.1016/j.aca.2017.11.044. Epub 2017 Nov 23.
Cell population based toxicity assays cannot distinguish responses of single cells and sub-cellular organelles; while single cell assays are limited by low statistical power due to small number of cells examined. This article reports a new single cell array based toxicity assay, in which cell responses at population level, single cell level and sub-cellular level can be obtained simultaneously at high throughput. The single cell array was produced by microcontact printing and selected area cell attachment, and exposed to damaging X-ray radiation, which was followed by fluorescence imaging after staining. Two image processing softwares written in Python and MATLAB were used to determine the expressions of proteins associated with cell migration and invasion, and production of reactive oxygen species (ROS), respectively. The results showed significant differences in responses at single cell level and distinctive molecular heterogeneity at sub-cellular level in a large population of cells upon exposure to radiation.
基于细胞群体的毒性检测方法无法区分单细胞和亚细胞细胞器的反应；而单细胞检测方法由于检测的细胞数量少，统计效力有限。本文报道了一种新的基于单细胞阵列的毒性检测方法，该方法可以高通量地同时获得群体水平、单细胞水平和亚细胞水平的细胞反应。单细胞阵列通过微接触印刷和选择区域细胞附着来制备，并暴露于破坏性的 X 射线辐射下，然后进行荧光成像染色。使用 Python 和 MATLAB 编写的两个图像处理软件分别用于确定与细胞迁移和侵袭以及活性氧 (ROS) 产生相关的蛋白的表达。结果表明，在暴露于辐射后，大量细胞在单细胞水平的反应存在显著差异，并且在亚细胞水平存在明显的分子异质性。
