Hello all-

I have tried to do a Masson's Trichrome stain for collagen on some
sections of rat heart which are horribly overfixed.  Staining was bad,
with weak collagen staining and a pale blue haze around portions of the
tissue.  I'm not sure whether the haze was due to the aniline blue
staining, or hematoxylin which didn't get rinsed out or something odd
like that.  In any case, we're trying to quantify the amount of collagen
present, but have trouble distinguishing true collagen staining from the
haze.

Is there any way to get better staining out of overfixed tissues?  Would
it help to just leave them in the aniline blue for an extended period of
time or perhaps alter the time in the bouin's step?  Any suggestions are
appreciated.

Thanks-

Brandon Schanbacher
--
Division of Pharmacology
Ohio State University