In situ High-Throughput Single-Cell Analysis Reveals the Crosstalk between Nanoparticle-Induced Cell Responses

Nanomaterials are widely used in a variety of industrial, biological, and medical applications. Therefore, high concerns about their possible impact on human and environmental health have been raised. Here, we describe a highthroughput single-cell imaging method to reveal the crosstalk among quantum dot (QDot)-induced ROS generation, apoptosis, and changes in nucleus size in macrophages. In triple marker combinations, we assessed the correlations of three QDot-induced cellular responses via divided subsets based on single-cell analysis. In contrast to the results obtained from the cell population, we demonstrated that the change in nucleus size was positively correlated with ROS generation. We found that QDot exposure induced ROS generation, which led to cell apoptosis, followed by a change in nucleus size. In general, these observations on crosstalk of cellular responses provide detailed insights into the heterogeneity of nanoparticle exposure.