[Histonet] Horse bones
gayle.callis <@t> bresnan.net
Wed Aug 6 10:42:25 CDT 2008
How big are the pieces? It would be advisable to have longer processing
schedule, rather than a routine schedule. This sounds like poor dehydration
and poor paraffin infiltration. Also, if your samples are less than 3 mm
thick, then you get the potato chip syndrome, where the bone is NOT
supported well enough, plus trimming should be done in very thin increments
(setting) with a SHARP blade. High profile blades work much better with
harder bone but if the bone is perfectly and totally fixed before
decalcification, tested, and processed properly, thin profile will work
although not as nicely.
You should do a decalcification endpoint test and there is a simple one that
works quite nicely, a weight gain/weight loss.
If the bone appears chalky or opaque, then the bone is either poorly
decalcified, poorly processed or both.
Longer infiltration in a harder paraffin i.e. Tissue Prep 2 ThermoScientific
ala Fisher is excellent for bone and three changes minimum.
The horse bone may be harder, depending on age of the animal, as compared to
Soaking only works IF the bone is well decalcified and processed. To avoid
surface decalcification, endpoint test. I will be happy to send the
protocol for that, but you need a balance that weighs in milligrams.
Gayle M. Callis
----- Original Message -----
From: <BERGERJL <@t> aol.com>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, August 05, 2008 6:07 PM
Subject: [Histonet] Horse bones
> We are trying to section pieces of horse femurs, fetlock and carpal
> They have been decalcified with either HCl decal or formic acid decal.
> Before routine processing the bones appear to be decalcified, but at
> they are very hard and brittle (even after surface decal) and chip out of
> paraffin block.
> Any suggestions would be helpful. A pathologist suggested a soap soaking
> solution but could not remember the name of the soap.
> Help and thank you,
> R. Berger, HT
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