[Histonet] uneven haematoxylin/eosin staining

Janice A Mahoney jmahoney <@t> alegent.org
Wed Jun 29 08:57:14 CDT 2005

I have experienced this problem at several labs where I have worked.  Finally figured out it happens more in the summer when the humidity is high.  It was due to incomplete removal of paraffin due to moisture in the xylene.  Try changing it more often or filtering it.  The filter paper will labsorbe the water.
Omaha NE

>>> Paul Bradbury <histology.bc <@t> shaw.ca> 06/28/2005 6:23:27 PM >>>
Sounds like a classical case of incomplete removal of wax before 
staining begins. Check the de-waxing reagents and times.

Paul Bradbury
Kamloops, BC

Diana McCaig wrote:

>We have been experiencing uneven staining recently.  Even when all sections
>are cut on the same microtome and put in the same rack on an autostainer,
>their macroscopic appearance show incredible variations.  It is so random we
>can not identify the problem.  The Harris hemotoxylin is filtered daily.
>The tissue type does not make any difference.  We can have 4 prostate slides
>and 2 will be good and 2 will show random staining intensities.  Recuts may
>show variation in a different area which makes me think it is the staining
>and not the cutting.
>Any suggestions would be truly appreciated.
>Diana McCaig, MLT 
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