[Histonet] holly nuclei bat man

[Rene J Buesa]

According with some tests I made and an article published in the JOH, the so called "nuclear holes" occur when a section with some remaining amount of water underneath is dried in an oven at 60ºC or more.
The "explanation" is that the heated underlying water "dissolves" nuclear material and hence the appearance of holes.
I could look for the reference of the JOH article if you want.
René J.

From: Patsy Ruegg <pruegg <@t> ihctech.net>
To: histonet <@t> lists.utsouthwestern.edu 
Sent: Saturday, January 5, 2013 3:37 PM
Subject: [Histonet] holly nuclei bat man

Happy New Year everyone!



Can we start a discussion once again on what causes holes in nuclei?  It is
pretty clear in my experience it has something to do with heating at least
for me with antigen retrieval.  When I do HIER at ph 8 especially I am
seeing a lot of holes in nuclei.  H&E on same tissue alone without HIER/IHC
doesn't seem to exhibit this artifact (holes in the nuclei).  I bet it can
also happen if the tissue is not fixed well enough to protect it from
paraffin processing but right now I am seeing it more after HIER, don't see
it as much on my IHC slides with no AR or when I use eier instead of hier.
Also notice that ph6 HIER and Ph9 HIER are not as bad as when I use Ph8 for
the same times and temps. 



Cheers,



Patsy



Patsy Ruegg, HT(ASCP)QIHC

Ruegg IHC Consulting, LLC

40864 Arkansas Ave

Bennett, CO 80102

Fax: 720-859-4110

<mailto:pruegg <@t> ihctech.net> pruegg <@t> ihctech.net



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