The fading after most retrieval procedures should not be very bad.  Heat
retrieval in citrate buffer causes very little nuclear fading in my tissues
(and no fading of lymphocyte nuclei).  Your staining and mounting procedure
may be the problem.  I find Vector's Nova Red compatible with (Harris)
hematoxylin.  Since Nova Red is insoluble in xylene, I can clear my tissue
and mount in resin (Permount) so that the nuclear staining is more easily
observed.

Allen A. Smith, Ph.D.
Professor of Anatomy
Barry University School of Graduate Medical Sciences
    Podiatric Medicine and Surgery
Miami Shores, Florida  33161


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Katri
Tuomala
Sent: Tuesday, April 04, 2006 7:06 PM
To: Guillermo Palao; Histonet
Subject: Re: [Histonet] Hematoxylin staining of frozen samples,where are the
nuclei?

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=2E 
> 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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