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A fluorescent molecule which is impacted by a light of a given wavelength is able to re-emit a light of higher wavelength. To characterize this fluorescence, it is possible to measure different parameters: fluorescence intensity, wavelength and lifetime, which represents the time during which the fluorescent molecule remains in its excited state. In lignocellulose, several molecules are fluorescent since they have aromatic groups, like lignin.

We have shown that the measurement of lifetime can demonstrate how lignin is organized and how it interacts with other cell wall polymers such as polysaccharides (cellulose and hemicelluloses): it thus can be related to lignocellulose recalcitrance. Also, fluorescence lifetime can even be measured for each pixel of an image, giving access to fluorescence lifetime images (see image above).

The objective of the book chapter presented here is to describe the protocol for preparing sample, measuring and analysing fluorescence lifetime data, so that it can be applied in different labs. This work was done in close collaboration between FARE and PICT platform of URCA.

Read: Terryn C, Paës G (2019). Fluorescence Lifetime Imaging of Plant Cell Walls in: Plant Cell Morphogenesis: Methods and Protocols, pp. 77-82 (Cvrčková, F. and Žárský, V., Eds.), Springer New York, New York, NY. https://doi.org/10.1007/978-1-4939-9469-4_5

Contact: Dr Gabriel Paës, gabriel.paes@inra.fr