Hi All,
I'm quite new at histology and am having a hard time getting good sirius red stains in my mouse livers. As you can see they're quite red and missing the yellow.
My protocol is:
1. Deparaffinise in xylene and 100-90-70% ethanol changes. Rehydate in ddH2O.
2. Stain with sirius red (Abcam AB150681)
3. Rinse in acetic acid solution twice
4. Rinse in absolute alcohol
5. Dehydrate in 2 changes of absolute alcohol
6. Mount
The slices are 5 micron.
Appreciate any advice!
You mean picro-siriusred, where siriusred stains connective tissue and picric acid stains cytoplasma.
The original protocol says 60 min incubation. Try to shorten the incubation. Try to prolong the rinse in acetic acid and check, if the red stain is well differentiated.
You work with FFPE tissue? Was the fixation and processing correct?
Dear Khali,
Which fixative did you use on your fragments?
Bouin's solution yields superior results for Sirius red staining.
Please make sure that all paraffin has been removed from the section before preparing it for staining.
Best regards
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