RE: EDTA pretreatment - sections falling of

From:"Sebree Linda A." <>

Hi Tibor,

Try your HIER procedure using EDTA in a 93-95 degrees C waterbath instead of
a microwave oven.  We've found that this is the gentlest protocol for
difficult sections such as fatty breast that tends to want to fall off.  We
simply bring our buffer and slides nearly to a boil in  a microwave oven
then transfer them to a waterbath for 10"-40".  For everything else that has
a HIER pretreatment, we use the Biocare Decloaking Chamber; it works great.
Another option you could try is substituting glycerol for the water in your
buffer; generally the higher boiling point of the glycerol prevents boiling
and is therefore gentler on your tissues.

Hope these recommendations are of value to you.

Linda A. Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
600 Highland Ave.
Madison, WI 53792-2472
FAX: (608)262-7174

-----Original Message-----
From: Tibor Ric []
Sent: Wednesday, February 21, 2001 11:53 PM
To: HistoNet Server
Subject: EDTA pretreatment - sections falling of

Hello Everybody!

We at our immunohistolab have started to use EDTA pretreatment (1mM; ph8,0;
4x5 min) for some antibodies. Our big problem is that  some sections
(expecially breast sections, cytospin materials and smears) all falling of
the glas...

Any suggestions to avoid the problem without chaning the buffert?

Thanks in advance!
Tibor Ric, Laboratory Scientist
Immunohisto lab at Karlstads hospital in Sweden.

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