- We identified four pixel classes in our images: green (leaves), purple (leaves and flowers), residue, and soil (Figure 1, left window).
- To facilitate sampling pixels from images, we wrote a macro for ImageJ that records x and y position and red, green and blue color density for each pixel selected (Figure 1) (link to macro - "Color Saver").
- We sampled the color of 10-15 pixels per class per image for 2-4 images from a given batch (same plant species, images acquired on the same day). We intentionally selected pixels to represent the full range of each of these color classes.
- Samples of pixels from each class revealed differences in coloration that could be used to classify pixels (Figures 2).
- We graphed the sampled pixels by class using various combinations of colors and color ratios. We found a broadly similar pattern for most of the images we sampled, though the exact threshold values varied depending on crop growth, lighting conditions and exposure, and camera.
Screenshot of pixel sampling in ImageJ using the "Color Saver" macro
Green (and some purple) pixels can be separated by low red:green ratio. They also typically have intermediate brightness. (Remember, we aren't separating gren from purple, but these 2 classes are different enough that they don't easily separate from the rest using a single rule).
The remaining purple pixels have a low green:blue ratio, and intermediate brightness.
We can separate out plant residue and soil based on red color and brightness; soil is generally quite dark, and residue quite bright. In many cases there is a region where these classes overlap. We consider this are to be ambiguous, and leave these pixels unassigned.