overfixed muscles

From:Barry Rittman <brittman@mail.db.uth.tmc.edu>

first I would rinse the tissues well after fixation to remove as much
formalin as possible.
You can attach the frozen sections in several ways.
1.    thin layer of gelatin placed on the coverslip and section allowed
to adhere to this. Slide then exposed to formalin fumes to fix the
gelatin. We used to use 1-2% gelatin. After all you have already
saturated the block with formalin so not much chance of overfixing with
formalin exposure of the sections.or
2.    Smear a thin layer of glycerin-albumen on coverslip. Then using
forceps gently heat the coverslip until fumes given off. Should do this
in a fume hood. I as told that  acrolein is formed from the glycerin and
partially fixes the albumen and also renders it stickier.
Hope that this helps

Chris Jones wrote:

> We routinely do a triple-fluorochrome stain on frozen sections of
> fixed
> laryngeal muscles. The 20u sections are cut onto gel-coated coverslips
> for staining. Recently we attempted this procedure on some muscles
> that
> had been left in formalin for almost two years, and these sections
> never
> stay affixed through the whole staining procedure. We also tried
> APTS-coated coverslips with the same result. Can this new problem be
> the
> result of overfixation? Any suggestions?

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