Using Genetically Encoded Fluorescent Biosensors for Quantitative In Vivo Imaging

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.