It is convenient to use Fiji’s color deconvolution plugin to do H-DAB (DAB-nickle) image analysis for the well stained IHC slides. But in the practical work, sometimes some slide’s DAB stain was not strong enough to be separated from the hematoxylin.  To create the specific vectors become necessary during the analysis.  Any suggestion for the detailed protocol to do that or could direct me to one? Is there alternative way to do the analysis other than creating the new vectors?

Thanks