Synopsis
Anisotropic, highly reflecting dye particles of microscopic size are visible in epithelial mucin stained with Alcian Blue. The formation of these exceptionally large dye deposits is attributed to the inherently high tendency of Alcian Blue to aggregate, together with the extremely porous and permeable character of hydrated mucin which allows the growth of dye crystalsin situ. Other basophilic tissue components may be too dense to permit a similar interstitial dye crystal growth.
Alcian Blue and many other basic dyes form reflecting deposits on both the natural and the cut surfaces of cartilage, and in the case of Alcian Blue much of the staining of cartilage in sections appears to be due to dye on the surface of, rather than inside, the matrix. The failure of Alcian Blue to penetrate deeply into most samples of cartilage was confirmed by block-staining experiments. It appears that at least two distinct mechanisms operate in Alcian Blue staining; this may account for some known properties of the dye, such as the relative ease with which strong salt solutions remove it from cartilage matrix.
Methods for distinguishing reflectance from fluorescence are discussed, and it is pointed out that reflectance would not be expected to occur with dye molecules in true solution, or molecularly dispersed through a stained substrate.
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Goldstein, D.J., Horobin, R.W. Surface staining of cartilage by Alcian Blue, with reference to the role of microscopic dye aggregates in histological staining. Histochem J 6, 175–184 (1974). https://doi.org/10.1007/BF01011805
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DOI: https://doi.org/10.1007/BF01011805