[Histonet] Water bath temperature

[Truscott, Tom]

Rebecca, Just a guess. Perhaps there's a reaction from the heat or water
or both that separates a component of the paraplast and makes an greasy
or insoluble bubble that blocks staining but disappears in the final
dehydration. Good Luck! Tom Truscott

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rebecca
Barnhart
Sent: Friday, February 11, 2005 11:24 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Water bath temperature

We were having a problem that the tissue would not take up the
hematoxlyin but would take up the eosin but only in some areas, so it
looked like polka dots.  We started one by one changing solution trying
to figure out where it was coming from.  We had all fresh solutions and
still had polka dots.  I tried the only other thing left to change, I
lowered the temperature of the water bath from 45 to 38 and this worked.
 We currently are using ParaPlast Xtra with a melting point of 52.  We
have all tried figure out why the temperature in the water bath will
make the difference.  Several times I have cut the same tissue at
different temperatures ranging from 38 - 45 and every time there are
polka dots at the higher temperatures.  Does anyone have any idea why
this would be?  

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