Using Genetically Encoded Fluorescent Biosensors for Quantitative In Vivo Imaging

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2200; p. 303
• Main Authors: Yoshinari, Akira, Moe-Lange, Jacob, Kleist, Thomas J, Cartwright, Heather N, Quint, David A, Ehrhardt, David W, Frommer, Wolf B, Nakamura, Masayoshi
• Format: Journal Article
• Language: English
• Published: United States 2021
• Subjects: Arabidopsis - genetics; Arabidopsis - metabolism; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Biosensing Techniques; Calcium - metabolism; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Optical Imaging; cpFP; Arabidopsis root perfusion chamber; FRET; Genetically encoded fluorescent biosensors; Fluorescence microscopy
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-0880-7_14

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 Ca in Arabidopsis thaliana using a ratiometric genetically encoded Ca indicator, MatryoshCaMP6s.