Re: sirius red for collagen
this is the same sirrus red protocol i use, except i do not store in fridge and it
lasts for more than 6 months.
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
> 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
> firstname.lastname@example.org 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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