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

LuAnn Anderson ander093 <@t> tc.umn.edu
Mon Nov 5 15:51:47 CST 2007

I too have had problems recently with Surgipath hematoxylin in brain 
tissue (not mucin). They replaced what I had with a new bottle, but I 
am seeing the same problems. I believe the formula has been changed 
to  eliminate mercury--don't know if this is the basis of the 
problems or not--but I believe that is when I started having 
problems. They have a "new" hematoxylin (560) and eosin (515), but 
our pathologists did not care for those either. I have been using 
Surgipath for 14 years and just started having staining problems with it.

LuAnn Anderson
Neuropthology Lab
University of Minnesota

At 03:08 PM 11/5/2007, Charles.Embrey wrote:
>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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