Movat's pentachrome stain?

Hi! I was wondering if anyone has any experience quantifying movat’s pentachrome stain using cellprofiler? I have a single .czi image (which i’ll split up into tiles) of which I want to measure the area of each colour in the stain. If anyone has a pipeline like this could you please share it? Cellprofiler version doesn’t matter.

Thanks in advance!

A staining method using up to 3 non-neutral colours could be separated via colour deconvolution. With 5 stains, the most difficult part would be resolving the co-localisation of the different dyes. Since histological stains follow a subtractive colour mixing model, one might end up seeing the same colour as a result of diifferent proportions of the 5 dyes.
If the dyes were “exclusive” (that is, no tissue component uptakes more than one dye) then HSB colour space could be a way forward, but I have no idea of how robust the assumption of exclusive staining is.

thanks for your insight! I suppose I could use an external script to segment the image using colour spaces prior to running cellprofiler to quantify the surface area. Or perhaps just the cellprofiler unmixcolours module.

Sure, you will need some sort of script to do anything beyond converting to some colour space, but that is a minor issue.
Take e.g. the colour green pixels appearing in some slides, that is (I guess) a colocolisation of alcian blue and Saffron, so if you try using HSB colour space, you have a problem on how to assess what is stained with alcian blue (blue) and what with the Saffron (yellow), because some tissue components are not exclusively stained by a single dye, they uptake several (e.g. cartilage, ends up green)… now add 3 more dyes to the mix. Pixels of a given colour might be due to different mixtures of the 5 dyes. It would be complicated to quantify what the colours mean (perhaps not possible if you are using RGB images). Maybe with hyperspectral imaging you can assess the absorption of each dye better, but I still think it might be quite complicated to do.
If you succeed, let us know!