Hi everyone! How do you standarize an image brightness relative to another image/images?
I was running into an issue while thresholding where the background for my images was different and throwing off what is considered signal or not. So my negative controls were coming up positive using the same threshold value as where other tissues signal was negative.
I have tried subtract background, but I need a way to standardize my images relative to each other as opposed to relative to itself. I hope that makes sense.
Thank you for the support!
The microscope I am using (NIKON Eclipse Ts2R Epifluorescence microscope) scales the background of the tissue based on the intensity of the signal, which has created a problem while I am quantifying. My negative control have no signal, but high background and I am unsure of how to accomodate that while I am quantifying. Therefore, while I am thresholding, my images are the software gives me false positive in majority of my tissues, while in others (such as Image 1 - Double Knockout CCN3) only high intensity signal is considered as threshold while everything else is not.WT%20TRITC%20GM|625x500