Figure 1.

Global procedure workflow. Plant stem transversal cross-sections are incubated with H*, G*, and S* monolignol reporters that become metabolically incorporated into the growing lignin polymer. Incorporated reporters are identified via a bioorthogonal chemistry strategy specifically linking each type of reporter to a different fluorescent probe allowing imaging by confocal microscopy (see Figure  2). Following image acquisition, a first representative image of the series is selected and the most suitable segmentation method (parametric or AI) is chosen and applied automatically to all acquired images. The data are then extracted for three different cell wall zones (CC, CML, or SCW) and relative reporter distribution and proportion determined by RM1 and RM2. Please see text for more details.