We also searched for a collagen staining with a good contrast for image
analysis. This is the protocol we use:

* biebrich scarlet-acid fushin solution (BS-AF)
   biebrich scarlet, aqueous 1% .... 90,0 ml
   acid fushin, aqueous 1%,......... 10,0 ml
   glacial acetic acid............... 1,0 ml

* phoshomolybdic-phosphotungstic acid solution (PMA-PTA)
   phoshomolybdic acid ................5,0 g
   phosphotungstic acid ...............5,0 g
   distilled water...................200,0 ml

* aniline blue solution (AB)
   aniline blue........................2,5 g
   glacial acetic acid.................2,0 ml
   distilled water....................100,0 ml

* 1% glacial acetic acid solution
   glacial acetic acid.................1,0 ml
   distilled water....................100,0 ml

staining procedure:
- deparaffinize and hydrate to distilled water
- BS/AF solution :3 X up and down
- rinse in distilled water
- PMA-PTA solution: 20 min
- AB solution: 8 min
- rinse in distilled water
- glacial acetic solution: 8 min
- dehydrate in alc 95%, isopropylalc, xyleen
- mount DPX

result:
collagen => blue
cytoplasm,keratin, muscle => red

you can also use weigert's iron hematoxyline (10min) for the nuclei, but for the
analysis it was better without for us.

good luck

Lobke De Bels


Department of Morphology
Faculty of Veterinary Medicine
Ghent University
Salisburylaan 133
9820 Merelbeke
Belgium
lobke.debels@ugent.be

Citeren Galina Deyneko :

> Dear Dr. Kiernan and Barbara, and colleagues,
>   I am also very interested in collagen staining using blue dyes for
> better contrast for image analysis. For now we use picro-sirius red
> overnight without counterstaining on mouse and rabbit hearts and arteries..
> Could you send me detailed protocol,
> please.I mean: time of incubation, wash, differentiation ect. As well could
> you give me a name of Dr. Kiernan book and where I can buy it.
>   Thank you at advance.
>   Galina Deyneko
>   Novartis Cambridge MA
>   617-871-7613
>
>
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