Most of the THCa signals that we have recorded appear to originate in structures with linear dimensions greater than 1 μm, as illustrated by the -stack series in Fig. 3. To create this image the laser focus was axially translated by 1 μm through the sample between individual frames. A total of 60 frames were recorded at a constant vibrational frequency of . Each frame was recorded in 10 s, including three averages. We verified that the bright signals correspond to a CARS signals by taking a hyperspectral image (not shown) at a slice in the center of the stack. To depict the three-dimensional structure of the entire trichome we have applied a -dependent color to each slice (from the -Stack functions of the MBF library in ImageJ). The hazy background in the top half of the image is the nonresonant background of the water immersion medium. Reds and yellows indicate regions toward the bottom of the stack, while blues and purples are at the top of the stack. There are four distinct regions that can be identified. The red/orange section at top right is highly structured and dense, and contains small voids or vacuoles. Directly beneath that section are two unconnected regions of similar morphology at different heights in the sample, colored yellow/green and light blue, respectively. These regions contain higher concentrations of cannabinoids, but in elongated and narrow structures. The left side of the trichome displays a large and more diffuse distribution without clear and sharp boundaries (yellow, green, light blue), similar to that observed in the bract. Finally, the far top right of the trichome has a number of very thin regions of cannabinoids, shown in green, that may be closely associated with or bound to cell membranes. From these images the compartmentalized distribution of the cannabinoids is readily apparent. The observation of these different morphologies has not, to the best of our knowledge, been previously reported.