Skip to main content

Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections


Sirius Red, a strong anionic dye, stains collagen by reacting, via its sulphonic acid groups, with basic groups present in the collagen molecule. The elongated dye molecules are attached to the collagen fibre in such a way that their long axes are parallel. This parallel relationship between dye and collagen results in an enhanced birefringency.

Examination of tissue sections from 15 species of vertebrates suggests that staining with Sirius Red, when combined with enhancement of birefringency, may be considered specific for collagen. An improved and modified method of staining with Sirius Red is presented.

This is a preview of subscription content, access via your institution.


  • Bancroft, J. D. &Stevens, A. (1977).Theory and Practice of Histological Techniques. pp. 129. Edinburgh: Churchill-Livingstone.

    Google Scholar 

  • Constantine, V. S. &Mowry, R. W. (1968). The selective staining of human dermal collagen. II. The use of Picrosirius Red F3BA with polarization microscopy.J. invest. Derm. 50, 419–23.

    Google Scholar 

  • Deitch, A. D. &Terney, J. Y. (1965). Effect of acetylation on acid dye binding and Sakaguchi reaction.J. Histochem. Cytochem. 13, 15–16.

    Google Scholar 

  • Ganter, P. &Jolles, G. (1969).Histochimie normale et pathologique. pp. 1517. Paris: Gautier Villars.

    Google Scholar 

  • Junqueira, L. C. U., Cossermelli, W. S. &Brentani, R. R. (1978). Differential staining of collagens type I, II and III by Sirius Red and polarization microscopy.Arch. histol. jap. 41, 267–74.

    Google Scholar 

  • Junqueira, L. C. U., Bignolas, G. & Brentani, R. R. (1979). A simple and sensitive method for the quantitative estimation of collagen.Anayt. Biochem. (in press).

  • Mueller-Eberhard, H. J. (1975). Complement.Ann. Rev. Biochem. 44, 697–723.

    Google Scholar 

  • Puchtler, H., Waldrop, F. S. &Valentine, L. S. (1973). Polarization microscopic studies of connective tissue stained with Picrosirius Red F3BA.Beitr. Path. 150, 174–87.

    Google Scholar 

  • Quintarelli, G. (1963). Masking action of basic proteins on sialic acid carboxyls in epithelial mucins.Experientia 19, 230–1.

    Google Scholar 

  • Spicer, S. S. (1962). Basic protein visualized histochemically in mucinous secretions.Exp. Cell Res. 28, 480–8.

    Google Scholar 

  • Stoward, P. J. (1975). A histochemical study of the apparent deamination of proteins by sodium hypochlorite.Histochemistry 45, 213–26.

    Google Scholar 

  • Sweat, F., Puchtler, H. &Rosenthal, S. I. (1964). Sirius Red F3BA as stain for connective tissue.Archs Pathology 78, 69–72.

    Google Scholar 

  • Tezuka, T. &Freedberg, I. M. (1972). Epidermal structural proteins. Isolation and purification of keratohyalin granules of newborn rat.Biochim. Biophys. Acta 261, 402–417.

    Google Scholar 

  • Vickerstaff, T. (1954).The Physical Chemistry of Dyeing. London: Oliver and Boyd.

    Google Scholar 

  • Weatherford, T. W. (1972). Staining of collagenous and non-collagen structures with Picrosirius Red F3BA.Ala. J. Med. Sci. 9, 383–8.

    Google Scholar 

Download references

Author information

Authors and Affiliations


Rights and permissions

Reprints and Permissions

About this article

Cite this article

Junqueira, L.C.U., Bignolas, G. & Brentani, R.R. Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem J 11, 447–455 (1979).

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI:


  • Collagen
  • Tissue Section
  • Collagen Fibre
  • Acid Group
  • Sulphonic Acid