I want to compare the above parameter for two samples. In one, cells are adherent and with full morphology on coverslip. While the others are non-adherent and round in shape. What I do is, using freehand selection tool of image j draw region around the total cell and again around the cytosolic portion of that cell and get the measure from the analyze tool. This is done for individual cells on the field for both the sample. Then I subtract the cytosol from the total mean to get the surface mean and then do the ratio. I want to know since, the shape of the cells for these samples are different whether it affects the mean value that I am getting? 2 images are being attached for convinience. Please help.