Using Genetically Encoded Fluorescent Biosensors for Quantitative In Vivo Imaging

Akira Yoshinari; Jacob Moe-Lange; Thomas J Kleist; Heather N Cartwright; David A Quint; David W Ehrhardt; Wolf B Frommer; Masayoshi Nakamura

doi:10.1007/978-1-0716-0880-7_14

Using Genetically Encoded Fluorescent Biosensors for Quantitative In Vivo Imaging

Methods Mol Biol. 2021:2200:303-322. doi: 10.1007/978-1-0716-0880-7_14.

Authors

Akira Yoshinari¹, Jacob Moe-Lange², Thomas J Kleist³, Heather N Cartwright⁴, David A Quint⁵, David W Ehrhardt^{2

4}, Wolf B Frommer^{1

3}, Masayoshi Nakamura⁶

Affiliations

¹ Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Nagoya, Japan.
² Department of Biology, Stanford University, Stanford, CA, USA.
³ Institute for Molecular Physiology, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany.
⁴ Department of Plant Biology, Carnegie Institution for Science, Stanford, CA, USA.
⁵ Center for Cellular and Biomolecular Machines (NSF-Crest), University of California, Merced, CA, USA.
⁶ Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Nagoya, Japan. mnakamu@itbm.nagoya-u.ac.jp.

PMID: 33175384
DOI: 10.1007/978-1-0716-0880-7_14

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.

Keywords: Arabidopsis root perfusion chamber; FRET; Fluorescence microscopy; Genetically encoded fluorescent biosensors; cpFP.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis / genetics
Arabidopsis / metabolism*
Arabidopsis Proteins / genetics
Arabidopsis Proteins / metabolism*
Biosensing Techniques*
Calcium / metabolism*
Luminescent Proteins / genetics
Luminescent Proteins / metabolism*
Optical Imaging*

Substances

Arabidopsis Proteins
Luminescent Proteins
Calcium