[Histonet] Decalcified Bone Clearing time

[Patsy Ruegg]

Make sure that you fix well in formalin before you decal (what are you using?), then I rinse off the decal solution (I use formic acid) and put them back into formalin before putting on the tissue processor for a longer than usual schedule and especially give them longer time to infiltrate in paraffin, at least 3 changes for 2 hours each after dehydration and clearing in xylene.

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting
40864 E Arkansas Ave
Bennett, CO 80102
H 303-644-4538
C 720-281-5406
pruegghm <@t> hotmail.com
pruegg <@t> ihctech.net


> From: WaitT <@t> livemail.uthscsa.edu
> To: histonet <@t> lists.utsouthwestern.edu
> Date: Mon, 17 Nov 2014 19:07:42 +0000
> Subject: [Histonet] Decalcified Bone Clearing time
> 
> Hey everyone! I'm having some issues about clearing the bone tissues after they get done dehydrating from decalcification.  I know that over clearing the bone tissues in Xylene for too long can cause them to become very brittle and I'm afraid that that was what was happening.  The bone blocks are not very big....their weights are listed as 0.222g, 0.100g, and 0.052g.  The biggest bone block is about 4mmx4mmx4mm so it's not very big I would say.  I cleared with 50% Ethanol/50% Xylene for 1 hour, and then I changed it with 100% Xylene for another hour.  The bone blocks become clear as expected but they begin to get a yellowish-tan tinge to them.  I'm not the most experienced but from what I've studied, I don't think that 2 hours in Xylene is overkill by any means so maybe it's something that I'm doing subconsciously.....if anyone has any suggestions are ideas please let me know! :)
> 
> 
> Trevor Jordan Wait
> University of Texas Health Science Center, San Antonio
> Class of 2017 MD Candidate
> Abilene Christian University Class of 2013 Graduate
> B.S.  Biochemistry
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