[Histonet] Decal question

[Patsy Ruegg]

Jackie I don't think the decal method would make much difference.  Lifting
of cortical bone and cartilage in my experience is the norm for bone
sections especially when you are trying to see bone integrated with soft
tissue (?your tumor cells I assume are soft).  The bone will pull away from
the cells.  What to do?  Well it does help to carefully drain the water off
the slides on edge when you pick them up and then dry them laying flat on a
heat plate at say 37dc overnight, then heat less than 60c, say 55c just hot
enough to melt the paraffin for several hours (?4)but with bone there is no
sure fire method to keep them intact that I know of except the old coating a
slide with 5% elmers glue let them airdry and use these slides to pick up
the section, but then if you are doing any kind of collagen IHC the glue
will interfer.  This is a great method for tintorial stains on bone sections
though.
If you are going to do HIER you are really going to be in for lifting of the
bone in my experience.  I use only enzyme digestion for bone IHC because I
can control it easier than HIER with less bone and cartilage lifting of the
section.
Patsy

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of
Jackie.O'Connor <@t> abbott.com
Sent: Wednesday, April 14, 2004 10:17 AM
To: Histonet (E-mail); histonet-bounces <@t> lists.utsouthwestern.edu
Subject: [Histonet] Decal question


I'm working on a trial with intra-tibial injections of tumors into mice.
Whole (skin and muscle trimmed) legs were fixed for about a month in 10%
NBF.  I used Surgipath's Decal I to decalcify (took about 6 hours - end
point determined by cutting through femur with scalpel).  Paraffin
sections cut nicely - I used a 48C waterbath to ensure no wrinkles, picked
up on positively charged slides.  I let them air dry overnight, then 60C
for an hour prior to H+E staining.  I'm not happy.  The tumors and
cortical bone are lifting and falling off the slides.  Any suggestions?
I've been reading old posts about EDTA decalcification - since hindsight
is 20/20 - would that have been a better choice??  Boo hoo.

Jackie O'
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