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Using Genetically Encoded Fluorescent Biosensors for Quantitative In Vivo Imaging

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Arabidopsis Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2200))

Abstract

Fluorescent biosensors are powerful tools for tracking analytes or cellular processes in live organisms and allowing visualization of the spatial and temporal dynamics of cellular regulators. Fluorescent protein (FP)-based biosensors are extensively employed due to their high selectivity and low invasiveness. A variety of FP-based biosensors have been engineered and applied in plant research to visualize dynamic changes in pH, redox state, concentration of molecules (ions, sugars, peptides, ATP, reactive oxygen species, and phytohormones), and activity of transporters. In this chapter, we briefly summarize reported uses of FP-based biosensors in planta and show simple methods to monitor the dynamics of intracellular Ca2+ in Arabidopsis thaliana using a ratiometric genetically encoded Ca2+ indicator, MatryoshCaMP6s.

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Acknowledgements

We thank Mr. Koji Matsushita (Nagoya University) for designing the perfusion chamber holder. This research was supported by the Human Frontier Science Program to M.N., JSPS Grant-in-Aid for Young Scientists (19K16164) to A.Y., and JSPS KAKENHI Grant (18KK0195) to M.N. and A.Y., and ITbM is supported by World Premier International Research Center Initiative (WPI), Japan.

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Authors and Affiliations

  1. Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Nagoya, Japan

    Akira Yoshinari, Wolf B. Frommer & Masayoshi Nakamura

  2. Department of Biology, Stanford University, Stanford, CA, USA

    Jacob Moe-Lange & David W. Ehrhardt

  3. Institute for Molecular Physiology, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany

    Thomas J. Kleist & Wolf B. Frommer

  4. Department of Plant Biology, Carnegie Institution for Science, Stanford, CA, USA

    Heather N. Cartwright & David W. Ehrhardt

  5. Center for Cellular and Biomolecular Machines (NSF-Crest), University of California, Merced, CA, USA

    David A. Quint

Authors
  1. Akira Yoshinari
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  2. Jacob Moe-Lange
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  3. Thomas J. Kleist
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  4. Heather N. Cartwright
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  5. David A. Quint
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  6. David W. Ehrhardt
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  7. Wolf B. Frommer
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  8. Masayoshi Nakamura
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Correspondence to Masayoshi Nakamura .

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Editors and Affiliations

  1. Centro Nacional de Biotecnología, CSIC Centro Nacional de Biotecnología, Madrid, Spain

    Jose J. Sanchez-Serrano

  2. Centro de Investigaciones Biologicas, CSIC Madrid Centro de Investigaciones Biologicas, Madrid, Spain

    Julio Salinas

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Yoshinari, A. et al. (2021). Using Genetically Encoded Fluorescent Biosensors for Quantitative In Vivo Imaging. In: Sanchez-Serrano, J.J., Salinas, J. (eds) Arabidopsis Protocols . Methods in Molecular Biology, vol 2200. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0880-7_14

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  • DOI: https://doi.org/10.1007/978-1-0716-0880-7_14

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