Hello, Thank you very much for your feedback. I am trying to become more familiar with the analysis but, unfortunately, not progressing as much as I want to.
First, I split the channels and only worked with the green and red channels, which correspond to my proteins of interest. I selected 3 ROIs in the image attached.
Unfortunately, I can not attach the image that I am working (ROIs) due to some error:"Maybe your image is corrupted." So, I screenshot it for your consideration.
I have been following the Coloc2 page and also the "Deconvolution is a great way to restore images to give a better estimate of the real fluorophore spatial distribution by suppressing noise and removing the offset or background..." section.
I tried to use the mask tool as well and select the correspondent description to further process the analysis, but all the times I receive the same warning message. Please, see attached:
I believe the issue must be on my end to be very unfamiliar with this type of analysis, but I am very thankful for your suggestions to overcome this difficulty.