[Histonet] DNA streaming out of nuclei in frozen sections?

[Rene J Buesa]

First I would determine if it is really DNA streaming out of the nuclei. Stain the sections with the Feulgen-Rossenbeck method, then I would also review the speed of freezing the tissue before FS, usually freezing time produce many artifacts in the final sections.
René J.


--- On Sun, 1/4/09, Pritchard, Michele <pritchm <@t> ccf.org> wrote:

From: Pritchard, Michele <pritchm <@t> ccf.org>
Subject: [Histonet] DNA streaming out of nuclei in frozen sections?
To: histonet <@t> lists.utsouthwestern.edu
Date: Sunday, January 4, 2009, 1:37 PM

Hello:  

I have recently begun to use frozen sections to localize an antigen found in
mouse liver tissue.  My previous experience has been almost exclusively with
staining of formalin-fixed, paraffin-embedded tissues.  I have done two runs of
this new staining protocol and noticed an odd artifact in both runs.  It appears
as though there are stringy stars throughout the sections!  The
'strings' seem to originate from the hepatocyte nuclei leading me to
believe that what I am seeing is DNA that has escaped the confines of the
nucleus and is streaming out over the tissue.  I did not squish or drag the
coverslips onto/over the sections; this is the only thing that I could imagine
would make such a mess.  Does anyone have any information on the nature of this
artifact and how to prevent it?  I will be happy to provide my protocol or a
picture if this is an odd artifact with which no one has experience.  I suspect,
however, that this is a 'newbie' mistake so I will refrain from posting
any additional information at the present time.

Thanks for your help.

---M.T. Pritchard

Michele T. Pritchard, Ph.D.
Research Associate
Nagy Laboratory
Department of Pathobiology/NE40
Lerner Research Institute
Cleveland Clinic
9500 Euclid Avenue
Cleveland, OH 44195
 
phone:  216.444.8613
fax:  216.636.1493
 
email:  pritchm <@t> ccf.org
 
Lab location:
Lerner Research Institute
NE4-214


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