RE: [Histonet] rabbit bone specimens

From:"Elizabeth Chlipala"

Mary
 
If the blocks are not decaled properly then it might help to soak the
blocks for about 20 minutes in decal solution and place them back on ice
and then cut them.  When we section any joints, after sectioning we
place the slides flat on a hot plate overnight.  The hot plate needs to
be around 45įC, warm enough for the sections to flatten out, but not too
hot that the paraffin melts. If they are too hot the articular cartilage
will curl.  One other thing, you need to let the slides drain a bit, any
water under the section when you lay it flat on the hot plate is going
to cause problems, but you canít let them drain too much so that the
sections dry out and turn white.  If you are staining with H&E you might
still get flipping of the articular cartilage, using a new sharp knife
can help with this. If you are staining with toluidine blue, this is
what you can do if your sections are fipping.  After the TB, rinse in
water, examine your slides individually for flipping and you can flip
the cartilage back by dipping the section gently in water to get the
cartilage to stay flat, then lay your slides flat on the hot plate to
dry and then coverslip from xylene. Good luck.  
 
Liz 
 
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz@premierlab.com
www.premierlab.com
 
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309
 
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mary
Lloyd
Sent: Thursday, December 16, 2004 9:25 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] rabbit bone specimens
 
I am working on a research projects with rabbit femurs and tibias.  They

are decalcified in cal-ex from fisher for 2 weeks and then processed for

1.5 hours in each solution on the processor.  2 changes of 70, 95, 100%,

xylene and then paraffin.  I am really having problems with the
cartilage 
folding right at the curve of the specimen.  It seems as though the bone

just below the cartilage is not thoroughly processed or  hard. When I do

feel I get a decent section it seems to fold when I stain.  I am using
plus 
slides. I don't normally do a lot of research so if anyone can help me
with 
this problem either with cutting or processing solutions I would really 
appreciate the help.
 
 
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