We have a project where the goal is to quantify Perls’ Prussian Blue Staining.
We have set a pipeline in QuPath to:
- set the stain vectors for color deconvolution;
- apply intensity based-thresholding to segment Perls’ staining;
- extract the area of Perls’ staining.
For now, we were extracting the Perls’ stained area as an output for the quantification, but we were wondering if we could also quantify the staining intensity.
I read that "*
Stain intensity quantification is meaningful only when dye uptake is stoichiometric (e.g. Feulgen reaction for DNA)
Landini, G., et al. (2020). “Colour Deconvolution – stain unmixing in histological imaging.” Bioinformatics.
Looking at the reaction formula for the detection of ferric iron in the Perls’ Prussian blue method, I think we could perform intensity quantifications.
Is it correct to perform intensity quantifications in Perls’ stained tissues?
If we could quantify the intensity of Perls’ staining, and taking into account the information available about Colour Deconvolution in Fiji should we use the 32-bit Absorbance output?
Thanks in advance,