[Histonet] baking bone slides for heat retrieval techniques?

[Patsy Ruegg]

There have been many shared methods for adhering bone sections to slides and
I have probably used them all at one time or another.  In my experience it
is best to avoid HIER on bone for IHC although I am sure many people do it
successfully.  I use enzyme digestion instead of heat retriveal for bone IHC
(either Proteinase K or Pepsin in most cases).  In my hands EIER is more
easily controlled and less harsh on tissues than HIER.  If I had to use HIER
on bone I would choose a more gentle method such as steam rather than
boiling.
My one cent worth.
Patsy Ruegg

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Garlits,
John
Sent: Tuesday, April 06, 2004 11:22 AM
To: HistoNet Server
Subject: [Histonet] baking bone slides for heat retrieval techniques?


I have tried repeatedly to use boiling or heated antigen retrieval
techniques on my bone slides, but the bone always peels off.  (The marrow
stays.)

I did hear somewhere that in this case one can heat the slides at around 55
degrees C overnight, which makes the bone adhere better to potentially
survive boiling antigen retrieval.  Has anyone tried this or know if this
works?

Thanks!

John Garlits, M.S.
Senior Research Technician
Hematology Oncology Division
Experimental Hematology Department
St. Jude Children's Research Hospital
332 N Lauderdale
Memphis, TN 38108
Phone: 901-495-3961
Fax: 901-495-2176


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