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Fluorescence microscopic distinction between elastin and collagen

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Summary

Distinction between elastin and collagen in arteriosclerotic lesions is difficult because immature and incompletely cross-linked collagen bind so-called elastica stains; furthermore, abnormal collagen can lack cross-striation and thus resemble elastin in electron microscopy. However, collagen and elastin differ significantly in their content of basic amino acids and hence in their affinity for heteropolyacids. This chemical difference was utilized for the development of a fluorescence microscopic method for distinction between collagen and elastin.

Paraffin sections of human autopsy material were treated with a 1% aqueous solution of phosphomolybdic acid (PMA) for five minutes, rinsed in distilled water, dehydrated and mounted. Other series were treated with the PMA-molybdenum blue reaction and with various special stains.

Only elastic membranes of aorta, the elastica interna and externa of sizable arteries, and “true” elastic fibers remained strongly fluorescent; the autofluorescence of collagen, reticulum fibers, basement membranes, pseudo-elastic fibers, and “elastic” membranes in small arteries was quenched. In other series PMA abolished the fluorescence of basic fluorochromes.

Correlation of fluorescence and direct light microscopic observations with chemical and electron microscopic data showed that the PMA-fluorescence method permits distinction between elastin and various types of collagen.

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  1. Department of Pathology, Medical College of Georgia, 30902, Augusta, Georgia, USA

    Holde Puchtler, Faye Sweat Waldrop & Linda S. Valentine

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  1. Holde Puchtler
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Puchtler, H., Waldrop, F.S. & Valentine, L.S. Fluorescence microscopic distinction between elastin and collagen. Histochemie 35, 17–30 (1973). https://doi.org/10.1007/BF00303661

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