RE: Picosirius Red stain for collagen

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From:"McCollough, Carol" <> (by way of histonet)
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Here is a picro-sirius red stain thread from Histonet.  Hope it helps.

Regards -
Carol B. McCollough, HT(ASCP)
Diagnostics & Histology Laboratory Manager
Maryland Department of Natural Resources
Fisheries Service
Cooperative Oxford Laboratory
904 S. Morris Street
Oxford, MD 21654

Subject: Sirius Red stain
From:    <> at INTERNET
Date:    2/9/98  11:31 PM

  I need some help recovering a stain for osteo components which one of my
pathologists swears is a "commonly-done" stain.  He thought it is called a
pico-sirius red...   Anyone know of a reference/recipe?

Sue Lewis
University of Iowa Hospitals and Clinics

From:           Patsy Ruegg <>
Subject:        Re: Sirius Red stain
Date:           Tuesday, February 10, 1998 10:21:30 EST

I use this stain for calcified bone.  It is 0.2% Sirrus Red made up in
saturated picric acid.  Stain sections for 20 min. then rinse several times
and let sit in dh20 for about 5 min. to remove the
yellow picric background.  This stain can be reused for 30 days and should
not be filtered.  It stains calcified bone red, nothing else stains.  It is
a good substitute for a silver von kossa stain.

From:           Gayle Callis <>
Subject:        picro-sirius stain
Date:           Tuesday, February 10, 1998  4:46:24 EST

Picro-sirius red is a used as a selective identification of collagen in bone
and soft tissue, and can be viewed with polarization.

Junqueira, LCU et all, Differential staining of collagens type I, II,, III
by sirius red and polarization microscopy,  Arch Histo Jap. vol 41, no 3,
1978.  pp267-274

Junqueira LCU et al, Picrosirius staiing plus polarization microscopy a
specific method for collagen detection in tissue section  Histochemical
Journal, 11 (1979) 447-455.  *** this article contains full method for

Junqueira, LCU et al.  Differential Histologic diagnosis of osteoid J
Pathology, v 148,(1986) 189-196.  *** This publication has  vivid and
beautiful color plates of stained tissues.

I had private communication with Junqueira about technic, anwashing after
the Picro-Sirius stain, he used running tap water wash until no color comes
out.  If your water is highly alkaline, he suggested using a slightly
acified water.  He also used fresh hematoxylin, no more than 2 months old
for best results.

Staining method was:

0.1% sirius red in saturated picric acid for 1 hour,
rapid wash in tap water
fresh Harris hematosylin for 6min.

Happy staining!

Gayle Callis

From:           Clint Lincoln <>
Subject:        Re: Sirius Red stain
Date:           Tuesday, February 10, 1998 13:33:36 EST

Hi Sue. Picro-Sirius red is a connective tissue stain. It does not stain
"BONE" per se but rather, it stains the bone collagen ( likely type I) which
forms first (coarsely fibered or woven bone commonly called osteoid)  and
then becomes mineral-laden (hydroxyapatite which is composed of calcium,
phosphate and hydroxyl ions). It was described very nicely by Holde Puchtler
in  "Manual of Published Histochemical Methods" by Holde in 1980. It is also
in Theory and
Practice of Histological Techniques by Bancroft and Stevens1996.

From:           Kevin Randall <>
Subject:        Re PSR
Date:           Thursday, October  2, 1997  3:32:55 EDT

Hi Mary

Our paediatric pathology lab routinely does elastic picro sirius red
instead of EVGs because one of the paediatric pathologists is colour blind
and finds EPSRs easier to interpret.

You can find our method at;

Hope this is of some help.



Kevin J. Randall
Dept of Histopathology
Queen's Medical Centre
University Hospital NHS Trust
Nottingham NG7 2UH
Tel: 0115 924 9924 x 43725

-----Original Message-----
From: Sharon Bledsoe []
Sent: Wednesday, November 17, 1999 4:10 PM
Subject: Picosirius Red stain for collagen

Does anyone have a procedure for picosirius red stain for collagen?


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