[Histonet] Water bath temperature[Scanned]

[Rebecca Barnhart]

Our stainer has an oven on it and it is set at 70.  And yes both slides
do go in the same oven.  To make sure everything was equal I stain both
slides at the same time.

>>> Kemlo Rogerson <Kemlo.Rogerson <@t> elht.nhs.uk> 2/14/2005 7:37:12 AM
>>>
After 'floating out' on the waterbath, do you dry in an oven afterwards
and,
if so, what is the temperature of the oven and do both sets (from the
45 and
38 waterbaths) go in the same oven for drying? For the same time?

Kemlo Rogerson
Cellular Pathology Manager
East Lancashire Hospitals NHS Trust
DD. 01254-294162
Mobile 0774-9754194
 

-----Original Message-----
From: Rebecca Barnhart [mailto:RBARNHART <@t> summithealth.org] 
Sent: 14 February 2005 12:21
To: histonet <@t> lists.utsouthwestern.edu 
Subject: RE: [Histonet] Water bath temperature[Scanned]

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 >>>
Rebecca;
I will only add a quick swiping question to the wiser replies you
should
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
differentiated
in 95% alcohol--- and got beautiful, consistent results. May I ask,
have
you removed the paraffin completely from the tissues before you tried
to
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
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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