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

Jennifer MacDonald JMacDonald <@t> mtsac.edu
Sun Nov 4 16:54:47 CST 2007

Have you always used the Gill Hematoxylin?  Mucin will stain with the Gill 

Jennifer MacDonald
Director, Histotechnician Training Program
Mt. San Antonio College
1100 N. Grand Ave.
Walnut, CA 91789
(909) 594-5611 ext. 4884
jmacdonald <@t> mtsac.edu

"DNon" <d.non <@t> verizon.net> 
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
11/03/2007 07:41 AM

<histonet <@t> lists.utsouthwestern.edu>

[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 
-  Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water 
-  .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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