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Abstract This work describes the development of a multi-inlet microfluidic device for in situ detection of multiple cytotoxicity endpoints. The device consists of an upstream microfluidic multi-inlet module (μMIM) and a downstream microfluidic cell culture channel (μCCC). The integration of a device with syringe pumps via the inlets of MIM mainly enables the performance of multiplex cytotoxicity assays targeted on multiple endpoints. The device in the study was used for the long-term culturing of BALB/3T3 fibroblast cells. In addition, cadmium ion was used as a toxicant to induce the toxicity in the cultured cells. The cytotoxicity endpoints, such as the in situ detection of reactive oxygen species (ROS), nuclear structures and cell morphology, were examined and correlated simultaneously for every single cell after they were exposed to cadmium. The data indicated that the percentage of cells that had produced ROS and shown nuclear staining were slightly higher than the cells that had produced ROS only, but this percentage was significantly higher than the cells that had shown only nuclear staining. The results revealed that the obtained correlated data from simultaneous observations of multiplex cytotoxicity assays were more efficient in providing the mechanistic explanation for Cd-induced toxicity than the individual data that were derived after examining a single endpoint. In addition, the results suggested that the intracellular formation of ROS induced by cadmium might have been followed by the alterations in cell and nuclear morphology and loss of membrane integrity. The device allows for the real-time monitoring of cells and their behavior against a toxicant and facilitates the rapid performance of cytotoxicity testing. Furthermore, we anticipate that the presented device could be very effective in advancing the understanding of the precise mechanism of cytotoxicity by improving the sensitivity and accuracy in identifying endpoints.
BioChip Journal – Springer Journals
Published: Mar 1, 2012
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