Cellular metabolites and ions can exhibit very specific spatiotemporal dynamics that are very challenging to monitor using extraction-based methods. Genetically encoded Föster resonance energy transfer sensors afford a powerful method of measuring these dynamics in situ and hence are now widely used in order to decode information communicated through the dynamics of cellular metabolites and ions. This methodology involves (1) the development of a suitable sensor, (2) genetic engineering of the sensor for its expression in the tissue of interest, and (3) measurement and characterization of the cellular metabolites and ions using optical imaging. This chapter describes the measurement aspects. We describe the imaging setup, sample preparation from leaf discs and root cells, performance of a perfusion experiment, and quantification of metabolite and ion concentrations from the imaging data. We also describe post-experiment analysis including estimation of sensor efficiency and spectral bleedthrough.
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 10 June 2014.
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