Re: intern seeks help...
|From:||Jeffrey S Crews <firstname.lastname@example.org> (by way of histonet)|
What is your fixative? In my experience with Masson's, we generally fixed
in NBF and then mordanted in Bouin's. Once we were given tissue fixed in
methanol, and it looked exactly like what you are describing: excessive
scarlet staining that could not be differentiated.
Also, what did you mean by "the control slides were coming out fine?"
What were those slides and where did you get them? If they are prepared
slides from another source, and are really coming out okay, then it would
indicate that there is something wrong with your tissue
Ihope this stirs up some ideas for you.
Jeffrey Crews, HTL (ASCP)
On Wed, 13 Sep 2000 09:20:25 -0700 (PDT) Heather Hall <email@example.com>
>My name is Heather Hall and I am presently doing an
>internship at Primedica Inc., located in Worcester,
>Mass. One of the required slides is skin stained with
>trichrome. For some reason, I cannot get this slide
>to come out right. I first tried Masson's procedure,
>with kit reagents. The slides appeared as though they
>had not been differentiated in
>phosphotungstic/phosphomolybdic acid long enough, as
>the collagen bundles were retaining the scarlet-acid
>fuchsin in their centers. My first step to correct
>the problem was to increase the time in the p/p acid;
>that didn't work. Next I made up fresh reagents;
>didn't work. Altering the times in the other reagents
>didn't work either, so I tried the Gomori's procedure.
> When the Gomori's did not work, I changed species
>(first species used was pig, second was human). The
>same problem was occuring in the human tissue also.
>Since the control slides appeared to coming out fine I
>tried a third species. I tried rat skin hoping that
>it might have a thinner connective tissue layer and
>permeability would be altered, but that came out as
>badly as the others. I don't know what to do next to
>try and fix this problem. Any suggestions would be
>very much appreciated. Thanks for any help you may be
>able to give.
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