[Histonet] uneven HE staining

Laurie Elmgren lelmgren <@t> sunriselab.com
Mon Jun 16 14:43:49 CDT 2008


Hi Paula and Atoska,

We have had the same problem from time to time. We have a Sakura DRS
200. I tend to think that the water was not fully drained from the
slides before racking them. Our procedure is to stand the slides up
vertically against the waterbath on paper toweling for at least 30
seconds before racking them. The water that remains under the tissue,
steams it in the oven, effecting the ability of the sections to take up
stain. We also change all of the solutions after every shift, and the
xylene and alcohol after ten full runs of slides. 
Laurie

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Atoska
Gentry
Sent: Monday, June 16, 2008 3:07 PM
To: Paula Wilder; Histonet
Subject: Re: [Histonet] uneven HE staining

Hello, when this type staining problem occurred in manual staining it 
was discovered to be caused by  incomplete deparaffinization.  
Corrective measure taken was to  replace clearing agents with fresh 
solution(s). However, I'm not familiar with the Tissue Tek Stainer or 
any of the other automated stainers. Best wishes. Atoska

Paula Wilder wrote:
> Hi everyone!
>  
> A colleague of mine is having uneven staining problems with their
automatic stainer, which I believe is a Tissue-Tek stainer.  This has
been an ongoing problem.  She tells me it is only on biopsies, very
intermittent.  On one slide she might have four sections; three stain
evenly and one does not.  The location of the unevenly stained section
is not always the same; sometimes it is the first section, sometimes the
last.  Several technicians are cutting so technique has been eliminated.
They tried tracking the biopsies to see if a particular doctor's office
was involved, but again, no. Does anyone have a clue or suggestion which
direction should be pursued next?  We don't think it is the processing;
dehydration?  I do not have their HE programmed times.
>  
> Thanks for your help,
> Paula Wilder
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