[Histonet] uneven haematoxylin/eosin staining

Geoff McAuliffe mcauliff <@t> umdnj.edu
Tue Jun 28 13:27:08 CDT 2005


In my experince incomplete removal of wax is a very common cause of 
uneven staining. I suggest fresh xylene (or whatever you use) and more 
changes, longer times, agitation or all of these.

Geoff

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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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029 
mcauliff <@t> umdnj.edu
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