Re: sirius red for collagen

From:rueggp

this is the same sirrus red protocol i use, except i do not store in fridge and it
lasts for more than 6 months.
patsy ruegg

Karen Bowden wrote:

> I used this on rat pineal tissue and thought is looked great.  The Investigator
> was expecting a lot less collagen so he didn't like it.  The collagen stain red
> and the other tissue stained yellow just like the "results" said it would.  It
> is also a very easy stain.
>
> Fix specimen in 10% Buffered Formalin, embed in paraffin, section at 5 microns.
>
> 1. Clearing agent                 -      5 min.
> 2. Clearing agent                 -      5 min.
> 3. Clearing agent                 -      5 min.
> 4. 100% Ethyl Alcohol          -      3 min.
> 5. 100% Ethyl Alcohol          -      3 min.
> 6. 95% Ethyl Alcohol            -      3 min.
> 7. 95% Ethyl Alcohol            -      3 min.
> 8. 80% Ethyl Alcohol            -     3 min.
> 9. Tap water wash               -     3 min.
> 5. 0.1% Sirius Red                -     30 min.
> 15. 80% Ethyl Alcohol          -     1 min.
> 16. 95% Ethyl Alcohol           -     1 min.
> 17. 95% Ethyl Alcohol          -     1 min.
> 18. 100% Ethyl Alcohol        -     10 min.
> 19. 100% Ethyl Alcohol        -     10 min.
> 20. Clearing agent               -     3 min.
> 21. Clearing agent               -     3 min.
> 22. Clearing agent               -     3 min.
> 23. Mount with mounting media
>
> Results:
>  light microscopy: collagenous fibers - red
>  other tissue elements - bright yellow
>
>  polarized microscopy:  collagen fibers - orange/red bands
>  against black background
>
> 0.1% Sirius Red Solution
> Sirius Red dye 0.1 gm.
> Picric Acid saturated  100 ml.
>
> Store in refrigerator.  Stable for 4-6 months.
>
> Reference:  Greenberg, Stephen, Ph.D., Laboratory Medicine, Vol. 17, No. 8,
> August, 1986.
>
> Karen Bowden
> University of CA, San Diego
>
> r-meyer2@northwestern.edu wrote:
>
> > Has anyone done a sirius red stain for collagen?  If so, could I please have
> > your protocol.  The AFIP histology manual shows a sirius red method for
> > amyloid.  But the researcher that gave me this project says he is not looking
> > for amyloid but collagen.  I mentioned trichrome but I guess he has already
> > tried this.  Thanks
> >
> > Bob Meyer, HTL
> > Northwestern University





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