Synopsis
Although collagen of either tendon or dermis can be stained equally well with Ponceau 2R/Acid Fuchsin or Light Green SF if the dyes are used independently, tendon collagen retains the red dye mixture and dermal collagen the green counterstain when the dyes are used sequentially in the Masson trichrome procedure. The results of experiments designed to assess differences in the penetration, retention and displacement of these arylmethane dyes have demonstrated that they are retained more firmly by the tensioned collagen of tendon or stretched dermis, and are more easily displaced from the collagen of relaxed tendon or dermis.
Experiments designed to test the basis of these differences in dye retention indicate that more positively-charged amino dye-binding sites are available in the tensioned collagen than in relaxed collagen, where they appear to be closely associated with adjacent carboxyl groups on the collagen fibres. The possibility that the carboxyl groups of associated acid mucopolysaccharides are implicated in the differences in staining propensity has been investigated and discounted. It is suggested that whereas the binding of arylmethane dyes to collagen under tension is through strong ionic linkages to amino groups, the binding of these and other dyes to relaxed collagen is through weaker hydrogen bonds. It is proposed that these differences in charge distribution on the collagen of the two situations is related to the previously described piezo-electric effect demonstrable on stretched collagen.
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Flint, M.H., Lyons, M.F., Meaney, M.F. et al. The Masson staining of collagen — an explanation of an apparent paradox. Histochem J 7, 529–546 (1975). https://doi.org/10.1007/BF01003791
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DOI: https://doi.org/10.1007/BF01003791