Guillermo,
Tissues go through a long procedure with immuno and the most detrimental for 
all tissue components is the heat retrieval and to a lesser degree enzyme=20
digestion. The less well fixed and processed tissue suffers most: connective 
tissue is distorted and nuclei stain very poorly.
Hope this explains your problem,
Katri

Katri Tuomala
Hamilton, Ontario, Canada

----- Original Message ----- 
From: "Guillermo Palao" 
To: "Histonet" 
Sent: Tuesday, April 04, 2006 5:22 PM
Subject: [Histonet] Hematoxylin staining of frozen samples,where are the 
nuclei?


> Dear histonetters,
>
>  When I stain samples placed on VWR frosted slides with Gills hematoxylin, 
> nuclei are nicely stained. However, when the same samples are 
> counterstained after passing through all necessary steps involved in 
> immunohistochemistry, hematoxylin staining of nuclei is simply horrible. 
> Is it possible that nuclei are lost in the different washing steps? Please 
> put forward any suggestions you might have.
>
>  Guillermo
>
>
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