Dear Yi,

The Picro-Sirius Red Stain Kit (Connective Tissue Stain) is intended for use in the histological visualization of collagen I and III fibers in addition to muscle in tissue sections. The PSR stain may be viewed using standard light microscopy or polarized light resulting in birefringence of the collagen fibers to distinguish between type I and type III.

http://www.abcam.com/picro-sirius-red-stain-kit-connective-tissue-stain-ab150681.html

Hoping this will be helpful,

Rafik

Dear Rafik, thank you for your comment. It is helpful.

http://stainsfile.info/StainsFile/stain/conektv/puchtler_psr.htm

but most interesting perhaps:  

RICH & WHITTAKER, 2005:

COLLAGEN AND PICROSIRIUS RED STAINING: A POLARIZED LIGHT ASSESSMENT  OF FIBRILLAR HUE AND SPATIAL DISTRIBUTION  

"Our goal was, using picrosirius red staining, circularly polarized light, and image-analysis software,

not only to identify fibers and quantify collagen content, but also to assess fiber hue and the spatial distribution  of the different colors.", cf: cf: jms.org.br/PDF/v22n2a06.pdf

or: OPEN ACCESS: VOGEL et al, 2015:

Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopy

https://www.ncbi.nlm.nih.gov/pubmed/26150980 or, more successful:  http://www.sciencedirect.com/science/article/pii/S2215016115000102

Best wishes and good luck! WM

PS: Note that there is also available a +/-  "special" staining sequence, namely SVG (Stevenel's Blue - Van Gieson stain).

What about type II collagen? Is there any method to stain for fluorescence microscopy?

I'm a bit confused, as the following study claims that picrosirius cannot differentiate between collagen types:

"As Junqueira and colleagues point out, many studies use color staining to differentiate collagen bundles and to specify collagen types, yet other studies report that polarized colors only reflect fiber thickness and packing. Using a simple histological example, our study illustrates the inability of Picrosirius red staining to differentiate collagen types, since the absorbed amount of polarized light by this dye strictly depends on the orientation of the collagen bundles." (Lattouf et al 2014 "Picrosirius red staining")

http://www.ncbi.nlm.nih.gov/pubmed/25023614

So, where do we stand? 

Thanks!

Dear Sophie Moittie,

thank you really for your reply to the thread, bringing up an interesting and important question, possibly with some consequences:

The article by Lattouf et al. 2014 [PDF: see: http://journals.sagepub.com/doi/pdf/10.1369/0022155414545787 ] states and concludes an "inability of Picrosirius Red staining to differentiate collagen types , esp. I & III"    (*)see below .

I have saved the article to my harddisk for further reading and perhaps will comment on later. 

(*) I admit that I had no possibility to test Picrosirius Red stain by myself and own tissue preparations using also polarizing microscopy.

But I know a bit about the ultrastructural appearance of these collagen fiber types and the difficulties to judge and interpret results of polarizing microscopical images. There are a lot of older AS WELL AS recent articles stating that discrimination of collagen types is possible, findings that perhaps can be disproved another time or by other experimental layout/design.

Interestingly, still suppliers of the dye (or staining kits) obviously are sure about the capabilities in distinguishing at least Coll. I &III: (only as ONE example:  cf:  http://www.abcam.com/Picro-Sirius-Red-Stain-Kit-Connective-Tissue-Stain-ab150681.pdf  or: cf: http://www.abcam.com/ps/products/150/ab150681/documents/ab150681-Picro-Sirius%20Stain%20Kit%20(website).pdf. DISCLAIMER: no affiliation with or financial  interest in any supplier or company (mentioned or not mentioned here).

I am exited to read any confirming or countering / different views from experts using Picrosirius Red in polarizing microscopy and having done in parallel IHC-staining specific for Collagen (fibres) type I, II and III.   Until then, I guess some people will use Picrosirius Red & polarizing microscopy to analyse Collagen types as recently: e.g.: 

 LC Junqueira et al, Arch Histol Jpn. 1978 Jun;41(3):267-74. Differential staining of collagens type I, II and III by Sirius Red and polarization microscopy. @  https://www.ncbi.nlm.nih.gov/pubmed/82432; 

CONTRARY to the publication of JUNQUEIRA 1979 (as cited in LATTOUF et al, 2014): Junqueira LC, Bignolas G, Brentani RR (1979). Picrosirius staining

plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem J 11:447-455.                                    

Z Haviarova et al - ‎2008: Comparison of collagen subtype I and III presence in varicose and non-varicose vein walls. @  https://www.ncbi.nlm.nih.gov/pubmed/18517131 

JM Street et al, ‎2014:  Automated quantification of renal fibrosis with Sirius Red and polarization contrast microscopy.                                                              @  https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4187565/ 

or  El-Deen Yassin et al, 2014:  Picrosirius red staining assessment of collagen after dermal roller application: A minimally invasive percutaneous collagen induction therapy.                                                                                                         @ http://www.ijdpdd.com/article.asp?issn=WKMP-0052;year=2014;volume=1;issue=2;spage=68;epage=74;aulast=Yassin

A matter of dispute therefore which might be elucidated by further practical reseach work. 

well explained below

http://www.ihcworld.com/_protocols/special_stains/sirius_red.htm

Based on Junquera et al 1979, a lot of works distinguish between collagen III and collagen I by their color after the use of picrosirius red F3BA dye and observation under polarized light. Red/yelow is associated to collagen type I and green to collagen type III, several authors such as Pierrard (1989), Rich and Withaker (2005) or Collins (2011) do not agree with this method of distinction between collagen types. Here is the arguments put forward by “Lattouf et al 2014”

(a) The first article of Junquera and its colleagues has been published before elucidation of heterotypic collagen fiber organization by Fleischmajer et al (1990) and Slepecky et al (1992). Thus, all collagen fibers are constituted by different types of collagens such as collagen type I, III, and V in vessels, skin, bone or gingiva or collagen type II and IX in cartilage. So, it seems to be difficult to distinguish, in the same fibers, between collagen type I or collagen type III after interaction with picrosirius dye. It also be noticed that the proportion of the different collagen types constituting collagen fiber regulate its diameter.

(b) Vascular Ehlers Danlos (vEDS) syndrome is characterized by COL3A1 mutations and lack of collagen type III in extracellular matrix. We use skin of patients suffering from this syndrome to show, as well as in skin of healthy subjects, the presence of greenish fibers after picrosirius red dying under polarized light. Lack of collagen type III in vEDS dermal extracellular matrix was checked by specific immunochemistry.

(c) After picrosirius red dying, we change by rotation of the microscopic stage, the angle between collagen fibers and the polarized light to demonstrated subsequent change of collagen fiber colors using normal healthy skins or collagen lattice sections.

So, if we concluded that Picrosirius red staining (under polarized light) should not be use to determine fibrillar collagen phenotypes, we remember that this dye is a powerful mean to measure physiopathological variations of collagen network in tissue sections associated to histomorphological analysis.

With my Best regards