I do believe this method is working, but I just have a feeling that something is still off about what I’m doing, plus there is very little consistency between images. ![]() Then Ctrl+M to get the %Area of what is within the ROI. And then I click add to place it into the ROI manager. Add threshold using more arbitrary numbers (33 for the top bar and 255 for the bottom). Subtract background (reducing to an arbitrary number for each image (50) which I have seen to show a lot of inconsistency and I hate this method). Close all of the windows except for the split channel that is labeled as “green”, which is actually grayscaled. I want to threshold the entire image but be able to adjust is accordingly so I can get a good representation of a percentage that is within the ROI. Here, you can see the original images plus my attempt at quantifying the percentage of thresholded amount of color within the ROI that I have drawn. “Segmentation in Fiji” workshop and corresponding slides. ![]()
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