Comparative analysis of colorimetric staining in skin using open‐source software

Colorimetric staining techniques such as immunohistochemistry ( IHC ), immunofluorescence ( IF ) and histochemistry ( HC ) provide useful information regarding the localization and relative amount of a molecule/substance in skin. We have developed a novel, straightforward method to assess colorimetric staining by combining features from two open‐source software programs. As a proof of principle, we demonstrate the utility of this approach by analysing changes in skin melanin deposition during the radiation‐induced tanning response of Yucatan mini‐pigs. This method includes a visualization step to validate the accuracy of colour selection before quantitation to ensure accuracy. The data show that this method is robust and will provide a means to obtain accurate comparative analyses of staining in IHC / IF / HC samples.