Hi Maria
Because of uneven shrinkage and the different rates at which tissues dry
on a warming plate, you will generally get some wrinkling ofparaffin
sections. What we used to do for cartilage, bone and teeth was to brush
out the wrinkles. (No I have not been at the Bourbon!)
Place the slide with most of the water drained on a slide warming plate.
Make sure that the paraffin wax  is not going to restrict the section in
the direction you are going to brush and that you do not have paraffin
smears as these will prevent the tissue from moving in that direction.
Using a fine sable brush gentle brush out the wrinkles. This is a skill
that needs to be acquired and depends on care in selecting the direction
of brushing and brushing out those tissues that dry fastest cartilage,
then bone, so you may screw up a few sections first. After brushing out
a small portion of the section, dry the brush by briefly touching to
some filter paper. The important thing is to go slowly and select the
best direction to brush. You are in effect gradually pushing the water
from under the section and at the same time are flattening the section,
removing the folds. This is superior to using needles as it will not
cause tears in the tissue. If you have not managed to brush out a
section before it starts to dry to much simply add a little fluid to the
fold and then try again. You have to be careful not to brush the soft
tissue as you will generally damage this.
Once section has been brushed and is dry then gently pass over a spirit
lamp to JUST melt the wax. This will aid in keeping the section on the
slide.
Two points
There are sometimes some folds that will simply not be removable and you
have to accept that.
Second is that if you have fluid on top of the section you will not be
able to remove folds from that area unless you can remove that fluid.
For those of you who are new to histotechnology, this method was
suggested by a trainee only two weeks after they started working with
us!
Call me if you need any further details.
Barry

713-500-4134


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Wadsworth, Marilyn P
Sent: Wednesday, January 28, 2004 8:52 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Articular cartilage wrinkles

Dear Histonet,

> I am processing specimens of articular cartilage/bone from rabbit 
> specimens.  The tissue has been fixed and decaled using EDTA in 
> formalin using the method of Kirviranta, 1985.
> 
> Kiviranta, I., et al., Microspectrophotometric quantitation of 
> glycosaminoglycans in articular cartilage sections stained with 
> Safranin O. Histochemistry., 1985. 82(3): p. 249-55.
> 
> The sections are paraffin embedded and sectioned at 5 or 6 microns. 
> Upon mounting on slides, the articular cartilage appears puckered or
> wrinkled (result of swelling of cartilage when exposed to water).   Is
> there anything that may be done to eliminate this artifact?  Is it 
> possible to float the microtomed sections on a non-aqueous solution? 
> I'd appreciate any tips you may offer.  Thanks
> 
Maria Roemhildt, PhD
University of Vermont
Department of Orthopaedics & Rehabilitation
95 Carrigan Drive
Burlington, VT  05405

Ph. 802-656-3823
fax:  802-656-4247

maria.roemhildt@med.uvm.edu


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