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

[DNon]

Smaller specimens are wrapped in Surgipath biowraps and placed in standard 
cassettes.  Larger specimens are sectioned and placed in standard cassettes.

We've never had problems with this protocol in the 10+ years I've been 
working here, but we're open to any suggestions.

----- Original Message ----- 
From: "Markus F. Meyenhofer" <micro <@t> superlink.net>
To: "DNon" <d.non <@t> verizon.net>
Sent: Saturday, November 03, 2007 1:40 PM
Subject: Re: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens


> In what capsules are your tissues processed??
> Regards,
> Markus F. Meyenhofer
> Microscopy Labs
> ----- Original Message ----- 
> From: "DNon" <d.non <@t> verizon.net>
> To: <histonet <@t> lists.utsouthwestern.edu>
> Sent: Saturday, November 03, 2007 10:41 AM
> 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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