[Histonet] (no subject)

[Cristi Stephenson]

I second Toni's suggestion.  Definitely sounds like the paraffin is still
present on the lower sections of the slide.

On Wed, Dec 4, 2013 at 10:17 AM, Rathborne, Toni <
trathborne <@t> somerset-healthcare.com> wrote:

> Could it be the heating/deparaffinization process? If the upper sections
> are staining more evenly, then maybe they are free from residual paraffin.
> Try extending the time in the ovens and/or xylene.
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:
> histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Chapman, Cherie J.
> Sent: Wednesday, December 04, 2013 1:12 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] (no subject)
>
> Hello all,
>
> I am looking for suggestions on issues with our H&E stain.
>
> I supervised a Veterinary Diagnostic lab for over 27 years and produced
> top quality sections, H&E's, special stains and IHC  on a variety of
> different species in our lab.
>  I am currently working in a  Dermatopathology Lab and I am  finding
> inconsistent staining with our H&E's. Working with just skin is a challenge
> all on its own.  We have made changes to our staining protocol and just not
> happy with the end product.
> What we are observing is inconsistent  staining on levels on the same
> slide.  The top section seems to stain more evenly than the middle and
> bottom sections.  I can actually see three different shades of color.   The
> specimens are ribboned sections so I know it is not from thick and thin
> sections.   We have looked at our processing times, microwave vs. oven
> times, staining reagents, different brands of hematoxylin and eosin,
> adjustments on staining times, tap water compared to distilled water.
>
> Our main processor is the Thermo Scientific STP-420 and our back up is the
>  Sakura VIP V processor.    I have been working with Thermo technical
> support thinking it might be a processing issue.  We have a Leica ST5020
> Multstainer/CV5030 Robotic Cover slipper we have made several changes that
> the technical teams has suggested to the reagents and staining time.  It's
> still not the quality that we are looking for.
> I have had culligan techs out several times to see if it could be
> something with  the water.
>
> We can run 100 slides the same day, same reagents and protocol and the H&E
> color is so inconsistent.
>  I would appreciate any suggestions in this matter.
>
>
> Cherie Chapman, BS, HT, HTL (ASCP)
> Associate Director of Dermatopathology Laboratory University of Missouri
> Department of Dermatology University Physicians Medical Building
> Phone: (573) 884-0123
> Fax: (573) 884-0834
>
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