Hi, I have been using ImageJ to measure the area of fetal ovaries, with Cleaved Caspase 3 and mvh/ddx4 expression for my IF. mvh is the green channel and CC3 is on the red channel.
I haven't had a problem measuring the area of expression of both of these individually (Colour>Split Channels>Adjust>Threshold>Create Selection>Measure, after setting scale and setting measurement as area).
What I need to know is, is there a way to measure the area that is expressing both red and green simultaneously? So that I can quantify the total area +ve expression of germ cell marker and cc3 in the ovary?
n.b. I have also counted the individual number of cells, but I don't think it is possible to manually count those that are CC3+, hense turning to area and density.
Give the Coloc 2 plugin a try: https://imagej.net/Coloc_2
https://imagej.net/Colocalization_Analysis
Melissa's suggestion is a good one and will give a lot better quantification then this:
Take your two masks and multiply them. This gives intersection. You can do measure or particle analysis after.
There is a way for quantification different colored labels adding a different program called Ilastik to the analysis process. I faced a similar problem while doing my work and it helped a lot, if you like feel free to reach out to me!
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