Abstract
In this protocol, we present a noninvasive in planta bioimaging technique for the analysis of hydrogen peroxide (H2O2) and glutathione redox potential in adult Arabidopsis thaliana plants. The technique is based on the use of stereo fluorescence microscopy to image A. thaliana plants expressing the two genetically encoded fluorescent sensors roGFP2-Orp1 and Grx1-roGFP2. We provide a detailed step-by-step protocol for performing low magnification imaging with mature plants grown in soil or hydroponic systems. This protocol aims to serve the scientific community by providing an accessible approach to noninvasive in planta bioimaging and data analysis.
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Acknowledgments
This work was carried out with the support of the NOLIMITS Center of Excellence for Plant Biology and Other Life Sciences established by the University of Milan. This study was carried out within the Agritech National Research Center and received funding from the European Union Next-GenerationEU (PIANO NAZIONALE DI RIPRESA E RESILIENZA (PNRR)—MISSIONE 4 COMPONENTE 2, INVESTIMENTO 1.4–D.D. 1032 17/06/2022, CN00000022). This study reflects only the authors’ views and opinions; neither the European Union nor the European Commission can be considered responsible for them. This work was also supported by a PhD fellowship from the University of Milan to S.B. and M.G., by the European Union’s HORIZON-TMA-MSCA-PF-E to E.D.A. (proposal number 101059695).
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Buratti, S. et al. (2024). Noninvasive In Planta Live Measurements of H2O2 and Glutathione Redox Potential with Fluorescent roGFPs-Based Sensors. In: Corpas, F.J., Palma, J.M. (eds) ROS Signaling in Plants . Methods in Molecular Biology, vol 2798. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3826-2_4
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DOI: https://doi.org/10.1007/978-1-0716-3826-2_4
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3825-5
Online ISBN: 978-1-0716-3826-2
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