[Histonet] sirius red
I have a question regarding sirius red staining.
Background: We are looking at amino acid based polyurethane 'mesh'
(fibrous structure) that are unseeded or seeded with porcine mitral valve
cells. The seeded scaffolds were either loaded or unloaded. We are
investigating the collagen production (we have used a pro-collagen assay
as well as a hydroxyproline assay and have come up with positive results).
In the past we have used Massons Trichrome to get some pretty pictures,
however, even using the room temperature mordent (overnight) was not good
for the polymer (biodegradable) so we have started to use sirius red
staining (thank you to the histonet members for the protocols we found on
Sections: samples were embedded in OCT. 5um sections were taken to be
There is one sample that we see staining (reddish)under bright-field and
polarized light (we are assuming this is collagen). For the majority of
the seeded scaffolds we are seeing 'cell-like' red staining which does not
show under polarized light (cytoplasm?). However, with the unseeded
controls we see similar (although not really cell-like) staining which is
predominently at/near the sample-OCT interface. Does anyone have any ideas
as to what it would be? I was thinking it could possibly be trapped dye
where the section has folded in slightly? The scaffold itself is staining
slightly pink too.
Thanks for your help
Department of Bioengineering
University of Washington
Seattle, WA 98195
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