Following laser induced changes of plant phenylpropanoids by Raman microscopy

Raman microscopy is a powerful imaging technique for biological materials providing information about chemistry in context with microstructure. A 532 nm laser is often used as excitation source, because high spatial resolution and signal intensity can be achieved. The latter can be controlled by laser power and integration time, whereby high power and long times give good signal to noise ratio. However, most biological materials absorb in the VIS range and fluorescence masking the signal or even sample degradation might be hindering. Here, we show that on lignified plant cell walls even very short integration times and low laser powers induce a change in the ratio of the lignin bands at 1660 and 1600 cm^-1. Time series on lignin model compounds revealed this change only in aromatic molecules with two OH-groups, such as coniferyl alcohol. Therefore, we conclude that monolignols are present in the cell wall and responsible for the observed effect. The solvent selectivity of the changes points to a laser induced polymerization process. The results emphasize how crucial careful adjustment of experimental parameters in Raman imaging of biological materials is and show the potential of time series and repeated imaging to get additional insights (e.g. monolignols).