[Histonet] cutting teeth

[Della Speranza, Vinnie]

Andi, you may want to contact Bob Skinner directly with your questions.

skinnerroberta <@t> uams.edu

I worked with Bob for about twelve years and he was and as far as I know is still using methyl salicylate in place of xylene as a clearing agent. Methyl salicylate avoids the brittleness and hardening that can occur with xylene.  I don't know if he's compared his clearing technique with Clear Rite. Bob's work is in orthopaedics, I don't know if he's done much with teeth recently but I know he did some eons ago.

Vinnie Della Speranza
Manager for Anatomic Pathology Services
Medical University of South Carolina
165 Ashley Avenue  Suite 309
Charleston, South Carolina 29425
Tel: (843) 792-6353
Fax: (843) 792-8974
 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Andrea Grantham
Sent: Tuesday, February 24, 2009 2:55 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] cutting teeth

I'm hoping that this isn't as painful as the actual process we all 
went through as children. I'm told that one of the investigators here 
will be sending a couple hundred teeth to my lab soon. I have never 
processed or sectioned teeth before so I'm hoping that I can find a 
few good suggestions on histonet.
Here is what they are doing - based on two papers from JOE in 05 and 07:
Fixation is in 10% NBF - how long do you fix teeth? Is temperature 
something to consider here?
Decalcification is done in a solution of equal parts of 50% formic 
acid and 20% sodium citrate. The papers say 10 days is enough to 
decalcify teeth. Is it enough time?
Following decal they infiltrated the teeth using "Skinner's method". 
Anybody know what this involves? One paper says Methyl salicylate was 
used as a clearing agent. Why is this better than traditional 
processing on my VIP using Clear Rite 3 as a clearing agent?
In the papers the teeth were embedded in Paraplast and sectioned at 5 
microns and then thinner, they actually want thinner sections - like 
2 microns. We are looking for pulp tissue.
Then they want a Brown and Brenn stain and maybe a trichrome.

Thanks,

Andi
.....................................................................
: Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
: Sr. Research Specialist       University of Arizona               :
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: (FAX:  520-626-2097)          (email:  algranth <@t> u.arizona.edu)       :
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