Re: TRAP

From:rueggp

we never had very good luck with NBF fixation for Trap stain.  i always used
cold absolute methanol for fixation and EDTA or no decalcification for TRAP
stain.  Cold (4dC) alcohol can be a good fixative maintaining good morphology
and enzyme activity, (the key here is that the fixative must be precooled and
kept cold for at least one hour) i know there have been articles using FFPE
tissue for TRAP stain but "in my limited experience with this method" it did
not consistantly work for me.
patsy

Nick_Madary@hgsi.com wrote:

> We are hoping to duplicate a TRAP procedure  following some fixationas and
> decals in some literature that one of our investigators read.  He had his
> techs do the fixaing/decal/staining all we did was cut.  They used the
> Sigma kit with varying results.  The investigator wants us to follow the
> procedure in the literature and it states that FFPE for 24 hr fix in NBF
> and then 24-48 hrs of decal in Surgipath HCL/EDTA decal.  The staining
> looks different when comparing the two sections with and without tartrate,
> but the pathologist says she does not believe it is staining the right
> stuff.  My belief is that the decal is too harsh, and maybe there is a
> better fixation method.  So now of course they want the "histotechs" to get
> involved(imagine that).  Any ideas on why this stuff may not be working?





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