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

[Cheri Miller]

We had the same issue and Richard Allen hematoxylin  solved it as well

Cheri Miller HT ASCP
Histology Supervisor
Physicians Laboratory Services, Inc.
Omaha, NE 68117
402 738 5052

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Saturday, November 03, 2007 10:59 AM
To: DNon; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens

I also had that very same problem and solved it by changing to Richard
Allan's modified Harris hematoxylin.
  René J.

DNon <d.non <@t> verizon.net> wrote:
  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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