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

DNon d.non <@t> verizon.net
Sat Nov 3 09:41:31 CDT 2007

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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