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Measure of Alkaline Phosphatase Activity of Plankton: An Integrated Microfluidic Approach

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Climate change is affecting the hydrodynamics of the world’s oceans and lead to an increase of the ocean stratification. This ocean stratification tends to trap phytoplankton in the nutrient depleted surface layer. To survive to this extreme environmental condition, some phytoplanktons can activate a set of enzymes (such as alkaline phosphatase) and use the dissolved organic matter as a nutrient source. Although measurement of the alkaline phosphatase activity can be performed with accuracy, no existing method can follow in real-time and at the single cell level the physiological adaptation such as the release of alkaline phosphatase by phytoplankton. As a consequence, no experimental evidence can directly link the alkaline phosphatase enzyme activity found in a sample to an individual. In this context, we plan to develop a new complete microfluidic system in order to better examine which species release alkaline phosphatase, in which extend and how the environmental conditions predicted in the future (Temperature and CO2) modify this enzymatic activity. This new microfluidic system will integrate in a same chip, a screening procedure based on the cell morphology, an incubation system, and an enzyme assay at a single cell level. Measurements of the kinetic of alkaline phosphatase will be performed by using two enzyme labelled fluorescence methods and time-lapse analyses.
Three main results are expected from this project. Firstly, the microfluidic device will be capable to encapsulate a wide spectrum of plankton species in single microenvironment. Secondly, the development of the automatic identification system based on the new image processing algorithm will create an easy method to classify planktons depending on their morphologies. This method could be purposed as valuable commercial application. The third technology developed in this study will be the creation of new tunable microenvironment capable to maintain target plankton alive during enzyme analyses.

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