[Histonet] Mucin overstained with Hematoxylin in G.I. specimens

[Bonnie Whitaker]

Hi All,

I had the same type incident (with the increase in mucin staining with the
Surgipath hematoxylin), but it was a one time thing that we attributed to a
larger than normal volume of H&E's in that particular time period.  We
increased the frequency that solutions are changed slightly, and haven't had
another problem.

Bonnie Whitaker


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Charles.Embrey
Sent: Monday, November 05, 2007 3:09 PM
To: DNon; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens


I find this post most interesting.  After using Surgipath Gills III for the
past 7 years we started having a problem with blue mucin just last month.  I
corrected the problem by adding GAA but I now have to monitor the pH on a
daily basis.  It is strange that after all these years I would suddenly have
this problem and then read about another lab having the same difficulty with
Surgipath Hematoxylin starting about the same timeframe.  Has Surgipath
changed something?

Charles Embrey Jr., PA(ASCP)
Histology Manager, Carle Clinic 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of DNon
Sent: Saturday, November 03, 2007 9:42 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens

Hello Histonet, 

     Over the past several weeks our lab has had problems with hematoxylin
overstaining in the mucin of our gastric specimens.  The degree of
hematoxylin overstaining is alarming in the mucin of the gastric specimens
but absent or negligible in other specimens.  One pathologist reviewing the
slides indicated some of the overstained specimens appeared to be
inadequately fixed.  We have addressed that issue which showed some
improvement, but the problem persists.  We are making changes to increase
fixation time (although the specimens now appear adequately fixed yet retain
a large measure of the overstaining), but any further suggestions for
corrective action would be much appreciated.

    We've been hesitant to increase acid alcohol clearing time because our
other specimens are staining well with the current protocol.

We are using the following reagents and stain times:

-  Surgipath specimen containers pre-filled with 10% neutral buffered
Formalin.
-  Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water
washes
-  .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash
-  Surgipath Scott's Tap Water for 1 min followed by running water wash

Sakura automated stainer set to agitate ( up and down dipping of racks )
once every 2 seconds, which is the fastest speed available on it.  


Dick Non
Pathology Department
Ocean County Medical Lab
Brick, New Jersey _______________________________________________
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