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An Innovative Continuous Flow System for Monitoring Heavy Metal Pollution in Water Using Transgenic Xenopus laevis Tadpoles

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posted on 01.12.2009, 00:00 by Jean-Baptiste Fini, Sophie Pallud-Mothré, Sébastien Le Mével, Karima Palmier, Christopher M. Havens, Matthieu Le Brun, Vincent Mataix, Gregory F. Lemkine, Barbara A. Demeneix, Nathalie Turque, Paul E. Johnson
While numerous detection methods exist for environmental heavy metal monitoring, easy-to-use technologies combining rapidity with in vivo measurements are lacking. Multiwell systems exploiting transgenic tadpoles are ideal but require time-consuming placement of individuals in wells. We developed a real-time flow-through system, based on Fountain Flow cytometry, which measures in situ contaminant-induced fluorescence in transgenic amphibian larvae immersed in water samples. The system maintains the advantages of transgenic amphibians, but requires minimal human intervention. Portable and self-contained, it allows on-site measurements. Optimization exploited a transgenic Xenopus laevis bearing a chimeric gene with metal responsive elements fused to eGFP. The transgene was selectively induced by 1 μM Zn2+. Using this tadpole we show the continuous flow method to be as rapid and sensitive as image analysis. Flow-through readings thus accelerate the overall process of data acquisition and render fluorescent monitoring of tadpoles suitable for on-site tracking of heavy metal pollution.

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