[Histonet] Water bath temperature

Rebecca Barnhart RBARNHART <@t> summithealth.org
Mon Feb 14 06:20:48 CST 2005

I am sure the paraffin is completely removed.  I know this because when
I stain a slide cut with the water bath at a temperature of 45 and at
the same time a slide cut with the water bath set at 38, the slide cut
at 45 will show polka dots and the slide cut at 38 will not.  

>>> "George Cole" <georgecole <@t> ev1.net> 2/13/2005 2:34:30 AM >>>
I will only add a quick swiping question to the wiser replies you
get---the fact that your tissue would not take up the hematoxylin is
interesting, because I stained thousands of my frozen muscle and nerve
section UNFIXED in hematoxylin and eosin--with the eosin
in 95% alcohol--- and got beautiful, consistent results. May I ask,
you removed the paraffin completely from the tissues before you tried
stain them? 
georgecole <@t> ev1.net 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu 
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
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
to figure out where it was coming from.  We had all fresh solutions
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
 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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