Real-time tracking of root hair nucleus morphodynamics using a microfluidic approach |
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Authors: | Gaurav Singh David Pereira Stéphanie Baudrey Elise Hoffmann Michael Ryckelynck Atef Asnacios Marie-Edith Chabouté |
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Affiliation: | 1. Institut de biologie moléculaire des plantes, CNRS, Université de Strasbourg, Strasbourg, 67084 France;2. Laboratoire Matière et Systèmes Complexes, UMR 7057, CNRS et Université de Paris, Paris, 75013 France;3. Université de Strasbourg, CNRS, Architecture et Réactivité de l'ARN, UPR 9002, Strasbourg, 67000 France |
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Abstract: | Root hairs (RHs) are tubular extensions of root epidermal cells that favour nutrient uptake and microbe interactions. RHs show a fast apical growth, constituting a unique single cell model system for analysing cellular morphodynamics. In this context, live cell imaging using microfluidics recently developed to analyze root development is appealing, although high-resolution imaging is still lacking to enable an investigation of the accurate spatiotemporal morphodynamics of organelles. Here, we provide a powerful coverslip based microfluidic device (CMD) that enables us to capture high resolution confocal imaging of Arabidopsis RH development with real-time monitoring of nuclear movement and shape changes. To validate the setup, we confirmed the typical RH growth rates and the mean nuclear positioning previously reported with classical methods. Moreover, to illustrate the possibilities offered by the CMD, we have compared the real-time variations in the circularity, area and aspect ratio of nuclei moving in growing and mature RHs. Interestingly, we observed higher aspect ratios in the nuclei of mature RHs, correlating with higher speeds of nuclear migration. This observation opens the way for further investigations of the effect of mechanical constraints on nuclear shape changes during RH growth and nuclear migration and its role in RH and plant development. |
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Keywords: | Arabidopsis thaliana live cell imaging microfluidics nucleus root hair single cell technical advance |
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