Making co-localized images

Hi everyone,

I’d like to create co-localized images from different colors by image J macro.

I have z-stacked tissue images with 2 fluorescent channels(EGFP and mCherry).
How can I extract only EGFP/mCherry double positive area?

By using coloc2 plugin, it seemed to be calculated statistical parameter about colocalization, but not to be created images.

Thank you,


Dear Tomohiro-gh,

the best approach is to generate binary masks for each channel by thresholding.
Image > Adjust > Auto Threshold…
To improve the segmentation you should use appropriate image processing (filters, background subtraction).

Then once you have the binary mask you can generate the intersection between the two using the image calculator:
Process > Image Calculator…
Using the multiply without the 32-bit option generates the result you want.


1 Like

Dear Christopher,

Thank you very much for your quick reply and great instruction.
I successfully created the image I wanted to!