Conventional and confocal fluorescence microscopy of collagen fibers in the heart

P. C. Dolber, M. S. Spach

    Research output: Contribution to journalArticlepeer-review

    140 Scopus citations


    The arrangement of collagen fibers has previously been studied with picrosirius red (PSR) staining and brightfield microscopy. We discovered that PSR staining can also be visualized by fluorescence microscopy. PSR-stained collagen was strongly fluorescent using excitation and barrier filters for rhodamine, and distracting background cytoplasmic fluorescence was drastically reduced with phosphomolybdic acid (PMA) treatment before PSR staining. The PMA-PSR fluorescence method was more sensitive than the brightfield PSR or PMA-PSR method, and permitted confocal microscopic study. We applied the method to the study of collagen fiber three-dimensional arrangement in perimysial and endomysial septa of the heart, showing the three-dimensional course of the fibers in stereo views generated by confocal microscopy. The PMA-PSR fluorescence method should be generally useful for accurately determining collagen fiber three-dimensional arrangement, a necessary prelude to mechanical modeling of collagen-reinforced tissues.

    Original languageEnglish (US)
    Pages (from-to)465-469
    Number of pages5
    JournalJournal of Histochemistry and Cytochemistry
    Issue number3
    StatePublished - 1993


    • Cardiac muscle
    • Collagen
    • Confocal microscopy
    • Dog
    • Extracellular matrix
    • Fluorescence microscopy
    • Heart
    • Picrosirius red

    ASJC Scopus subject areas

    • Anatomy
    • Histology


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