I have an H&E stained image. For the residual staining (stain 3), the red and green values are negative. Is this normal or do I need to re-estimate the staining vectors?
That is fairly normal in areas where another stain has high intensity. It will tend to become more severe as you have a higher Background set.
It happens with other stains too, for example you might see negative hematoxylin intensities where DAB staining is very dark or “perfectly DAB.”
Thank you! I’m glad you’re here on this forum. You’ve been very, very helpful!