Re: sirius red for collagen
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,
University of CA, San Diego
> 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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