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Method for Picrosirius Red-Polarization Detection of Collagen Fibers in Tissue Sections

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Fibrosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1627))

Abstract

The extracellular matrix is critical in guiding cell behavior in normal and pathologic tissues, particularly in cancers and fibrosis. Highlighting the organization (or disorganization) of a collagen-containing matrix can be very useful for understanding or grading pathophysiological conditions. The picrosirius red stain (also called “Sirius red” stain) is one of the best understood histochemical techniques able to selectively highlight collagen networks. Relatively inexpensive, the technique relies on the birefringent properties of collagen molecules. While the picrosirius red stain alone does not selectively bind collagen network, it becomes more specific than the other common collagen stains when combined with polarized light detection. This is why the selective histochemical procedure for collagen detection should be called the picrosirius-polarization method. In this chapter, we will provide essential explanation and detailed protocols and tips to allow collagen researchers not only to better understand how this staining technique works but also to easily apply this technique to their collagen-related research.

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References

  1. Levame M, Meyer F (1986) Le Picropolychrome de Herovici: application à l'identification des collagènes de types I et III. [Herovici's picropolychromium. Application to the identification of type I and III collagens]. Pathol Biol 35(8):1183–1188

    Google Scholar 

  2. Junqueira LC, Bignolas G, Brentani RR (1979) Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem J 11(4):447–455. doi:10.1007/BF01002772

    Article  CAS  PubMed  Google Scholar 

  3. Junquiera LC, Junqueira LC, Brentani RR (1979) A simple and sensitive method for the quantitative estimation of collagen. Anal Biochem 94(1):96–99. doi:10.1016/0003-2697(79)90795-4

    Article  CAS  PubMed  Google Scholar 

  4. Muller-Eberhard HJ (1975) Complement. Annu Rev Biochem 44:697–724. doi:10.1146/annurev.bi.44.070175.003405

    Article  CAS  PubMed  Google Scholar 

  5. Acton S, Resnick D, Freeman M et al (1993) The collagenous domains of macrophage scavenger receptors and complement component C1q mediate their similar, but not identical, binding specificities for polyanionic ligands. J Biol Chem 268(5):3530–3537

    Google Scholar 

  6. Vogel B, Siebert H, Hofmann U et al (2015) Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopy. MethodsX 2:124–134. doi:10.1016/j.mex.2015.02.007

  7. Orringer JS, Rittié L, Baker D et al (2010) Molecular mechanisms of nonablative fractionated laser resurfacing. Br J Dermatol 163(4):757–768. doi:10.1111/j.1365-2133.2010.09998.x

  8. Drifka CR, Loeffler AG, Mathewson K et al (2016) Comparison of picrosirius red staining with second harmonic generation imaging for the quantification of clinically relevant collagen fiber features in histopathology samples. J Histochem Cytochem 64(9):519–529. doi:10.1369/0022155416659249

  9. Whittaker P, Kloner RA, Boughner DR et al (1994) Quantitative assessment of myocardial collagen with picrosirius red staining and circularly polarized light. Basic Res Cardiol 89(5):397–410. doi:10.1007/BF00788278

  10. Bromage TG, Goldman HM, McFarlin SC et al (2003) Circularly polarized light standards for investigations of collagen fiber orientation in bone. Anat Rec B New Anat 274(1):157–168. doi:10.1002/ar.b.10031

  11. Rich L, Whittaker P (2005) Collagen and picrosirius red staining: a polarized light assessment of fibrillar hue and spatial distribution. Braz J Morphol Sci 22(2):97–104

    Google Scholar 

  12. Borges LF, Gutierrez PS, Marana HR et al (2007) Picrosirius-polarization staining method as an efficient histopathological tool for collagenolysis detection in vesical prolapse lesions. Micron 38(6):580–583. doi:10.1016/j.micron.2006.10.005

  13. Nerurkar NL, Baker BM, Sen S et al (2009) Nanofibrous biologic laminates replicate the form and function of the annulus fibrosus. Nat Mater 8(12):986–992. doi:10.1038/nmat2558

  14. Gadd VL (2014) Combining immunodetection with histochemical techniques: the effect of heat-induced antigen retrieval on picro-Sirius red staining. J Histochem Cytochem 62(12):902–906. doi:10.1369/0022155414553667

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Correspondence to Laure Rittié .

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Rittié, L. (2017). Method for Picrosirius Red-Polarization Detection of Collagen Fibers in Tissue Sections. In: Rittié, L. (eds) Fibrosis. Methods in Molecular Biology, vol 1627. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7113-8_26

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  • DOI: https://doi.org/10.1007/978-1-4939-7113-8_26

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7112-1

  • Online ISBN: 978-1-4939-7113-8

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