Re: [Histonet] Re: Processing fatty breast tissue and fatty derm tissue

From:Rene J Buesa



Anthony:
  Why would you after dehydrating and starting the clearing stage would go back to a dehydrating step. I don't see any rationale in doing that.
  The thing with fatty tissue is not that they need additional dehydration (since fatty tissue contains less water that other tissues) but they need a complete elimination of fat obtained with the clearing agents.
  What I used to do was to eliminate the las 2 dehydrating steps, and substitute them with one half:half with dehydrant:clearing followed by an additional clearing and it always worked very well for me.
René J.
anthony@histotechexchange.com wrote:
  Dear Ramona:

I am late to the game so I am sorry if you have already had this one.

I have never done this, so please try it on some fatty tissue that you are
going to throw away and secondly I have no idea how it will affect antigen
retrieval or anything involving antibodies.

With that out of the way you may want to try taking the specimen through
alcohols then onto the first clearing agent, then back to alcohol and then
back into the clearing agents and on to wax.
Good luck on finding the times etc.

Can anyone else out there give your opinion on this practice.

Yours truly,

Anthony Williams BSc. HT
Histotech Exchange LLC
19 Whitmore St.
Lexington, VA 24450
T 1 (302) 383 9780
F 1 (540) 463 3583
anthony@histotechexchange.com




Dear Ramona:

I am late to the game so I am sorry if you have already had this one.

I have neve done this, so please try it on some lipoma that you are going
to throw away and secondly I have no idea how it will affect antigen
retreval or anthing involving antibodies.
> I also would like some advice on processing fatty tissue. I have tried
> using Pen-Fix prior to processing, but my pathologist complain about an
> artifact and refuse to use it anymore. Can't explain that one. I am
> currently processing with formalin x2, 70% x2, 95% x2, 100% x2, clearing
> x2. I thought the dehydration step should be increased on the fatty
> tissue, but you are saying that is not the problem? Can someone else
> please offer some experience on this issue.
>
> Ramona Turner, HT (ASCP)
> Potomac Hospital
> Woodbridge, VA
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