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

DNon d.non <@t> verizon.net
Mon Nov 5 17:51:18 CST 2007

I spoke with Surgipath's QC department today and asked if there was a change 
in their Gill's III formulation.  According to Mike Urban at Surgipath, 
nothing has changed.  Nonetheless, he is going to run additional tests on 
the lot number in question and will get back to me.  I find your post very 
interesting indeed as our lab has used Surgipath Gill's III for the last 10 

They ( Surgipath) will also be sending me a gallon of Gill's III from a 
different lot.  If it doesn't show improvement, we'll either lower the ph 
similar to what you did, or switch to Harris hematoxylin.

----- Original Message ----- 
From: "Charles.Embrey" <Charles.Embrey <@t> carle.com>
To: "DNon" <d.non <@t> verizon.net>; <histonet <@t> lists.utsouthwestern.edu>
Sent: Monday, November 05, 2007 4:08 PM
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

    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
-  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
-  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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