[Histonet] Weight Loss/Weight Gain Decal

[Jennifer MacDonald]

I believe this was originally from Patsy Ruegg

Decalcification End Point: Weight Loss, Weight Gain


1.      Blot sample to remove excess fixative
2.      Weigh bone in mg, record as beginning weight
3.      Next day, rinse bone, blot and weigh bone daily, record weight. 
Change decalcifying solution to refresh acid OR EDTA. Return bone to fresh 
decalcifying solution.
4.      When bone begins to GAIN weight, remove from decalcifying 
solution, rinse and process.

        YOU MUST WEIGH IN MILLIGRAMS FOR ACCURACY

Once calcium is totally removed (bone loses weight as this happens), water 
replaces the calcium and weight begins to go up.  This is the point at 
which calcium should be totally gone.  The original method used a chemical 
test at the end to insure no calcium was in the decalcifying solution.  If 
you do this, you cannot stir the solution during decalcification.  Be sure 
to suspend bone in the solution to insure all sides of bone are in contact 
with decalcification solution. 



From:   "Wait, Trevor Jordan" <WaitT <@t> livemail.uthscsa.edu>
To:     "histonet <@t> lists.utsouthwestern.edu" 
<histonet <@t> lists.utsouthwestern.edu>
Date:   08/11/2014 12:29 PM
Subject:        [Histonet] Weight Loss/Weight Gain Decal
Sent by:        histonet-bounces <@t> lists.utsouthwestern.edu



Hello all! I'm currently doing some decalcification and was curious if 
anyone had some particular advice about the weight loss/weight gain 
method. I understand that when the decalcification process is complete, 
the tissue block will begin to increase in weight. However, I'm confused 
when I should record the weight for the block once they have been taken 
out of the EDTA solution. You see, for the times I weighed the blocks 
before... the weights were a little skewed because there were differing 
amounts of solution on the blocks while they were sitting on the balance. 
I just want to standardize my protocol a little more so that I can be sure 
the block is actually gaining weight due to the calcium loss rather than 
just extra solution sitting on the outside of the block. Would letting the 
blocks sit out of solution for about 30 minutes before being weighed help 
with the matter? I know that the blocks take on water once they are 
completely decalcified so I'm not sure how much this will affect that.


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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