I am looking for a way to calculate the intensity ratio out of two ROIs within one image (see exemplary image below). As my goal will be to calculate the polarization anisotropy out of those two ROIs, I moreover don’t want to calculate only a usual ratio like r=I1/I2 but rather a modified ratio like r=(I1-I2)/(I1+2*I2) with I1 and I2 being the pixel intensities of both ROIs. As I wish to do this while live recording using MicroManager, I want to avoid to just duplicate both ROIs as single images and doing the calculation afterwards.
So far, I’ve tried using the Ratio Plus plugin but this seems to not be able to calculate a ratio out of two ROIs instead of out of two images. Also, I couldn’t find a possibilty to modify the ratio-equation the way I wish to.
I hope any of you might have some helpful tips on this issue.