2013), as it is essentially a two-dimensional procedure ( Mandarim-de-Lacerda 2003 Marcos et al. Morphometric image analysis is highly influenced by threshold settings, magnification and image resolution ( Huang et al. (2014), the authors state that picrosirius red associated with morphometric image analysis “remains the most powerful method to study and quantify collagen” and we would like to comment on this sentence. This is well explained by the findings presented by Lattouf et al. 2011) or with Masson’s trichrome ( Street et al. Notably, the assessment of collagen area with picrosirius red and polarization has been weakly correlated with the area of collagen III detected by immunohistochemistry ( Farris et al. Picrosirius red is considered ideal for collagen staining because it does not fade, is selective and is highly reproducible, more so than Masson’s trichrome or collagen immunohistochemistry ( Farris et al.
Nowadays, this stain is widely used for studying fibrosis in many organs and, particularly, in the liver (e.g., Huang et al. Using this approach, the authors definitively clarified a frequent misconception with regard to the use of this stain, showing that picrosirius red does not allow a differentiation of collagen types in polarized microscopy. (2014) combined polarization microscopy, immunohistochemistry and connective tissue reconstruction to study the collagen staining of picrosirius red. In a recent issue of this journal, Lattouf et al.
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