SRS images and scanning electron micrographs of the cuticle of T. salsuginea following cold-induced wax biogenesis. T. salsuginea leaves were imaged using SRS spectral imaging and SEM before and after cold treatment. A, C, E, and G, Three-dimensional reconstructions of image stacks from a 250- × 250-µm field of view at the 2,840 cm–1 Raman shift. B, D, F, and H, SEM images. A to D are, respectively, from the adaxial and abaxial leaf surfaces of plants grown at 20°C for 8 weeks. E to H are, respectively, from adaxial and abaxial leaf surfaces of plants grown at 20°C for 4 weeks and then at 4°C for 2 weeks and at 20°C for an additional 2 weeks. Bars = 50 µm.