[Histonet] processing problem

[Barry Rittman]

Hi Martha
I think that Caroline is correct but you have nothing to lose by removing
the  wax and then trying to dehydrate and so on. A more gentle way after
removing the wax and dehydrating is to use chloroform instead of xylene and
hand processing. Chloroform is much more gentle although tissue do usually
float in it an do not become transparent. However unlike xylene, tissue can
be left in chloroform for hours to overnight without any appreciable
hardening.  Then after chloroform you can leave the tissues in a mixture of
chloroform/wax (1:1) at room temperature again for hours to even overnight.
During this process some wax does penetrate into the  tissue and therefore
can leave in wax for less time with less chance of hardening.
Yours is certainly not the first lab that this has happened to.
Good luck to you.
Barry

On Mon, Feb 16, 2015 at 11:30 AM, Caroline Miller <mills <@t> 3scan.com> wrote:

> I am afraid your tissue might be 'toast'. I tried to reprocess from that
> same issue and things did not go well. We were processing mouse tissue,
> which is dry enough, but then when we put the tissues back through xylene
> and into 100% and back again it got even worse.
>
> We had to just get what we could from the blocks and confess to the
> researchers what had happened :(
>
> sorry for the bad news, I hope someone has better for you!
>
> mills
>
> On Thu, Feb 12, 2015 at 5:41 AM, Davenport, Martha R <
> martha.davenport <@t> uky.edu> wrote:
>
> > I am Martha Davenport, supervisor, at the University of KY histology lab.
> > We had a processing problem caused  by  accidently placing 70% (instead
> of
> > 100%) into the last dehydration container. Would anyone please give me
> info
> > on how you would remedy this?  We usually reprocess the tissue but have
> had
> > trouble in the past with the tissue morphology being optimal. Any help
> will
> > be greatly appreciated.
> > Martha Davenport 859-257-1822
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>
>
>
> --
> Caroline Miller
> Director of Histology
> 3Scan.com
> 415 2187297
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