Single cell patterning for high throughput sub-cellular toxicity assay
Abstract 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. Highlights Single cell array is made by selective cell adhesion onto lithographically patterned surface. Population based toxicity, single cell toxicity, and subcellular toxicity are derived simultaneously at high throughput. Issues of cell overlapping and clustering are completely solved. Imaging software allowed for rapid, objective and automated extraction of toxicity information without user interference. Graphical abstract [DISPLAY OMISSION]
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