[Histonet] RDO question

[Kemlo Rogerson]

So why not use EDTA? That can decalcify very dense bone including teeth. Try
5 to 10% at pH 6.5 to 7.5 to prevent protein hydrolysis. 

You could try 1M Trichloroacetic acid, nitric acid, formic acid, Picric acid
OR EDTA at raised temperatures (26, 29, 30 or 40 degrees C) with alternating
current (50 mA, 100 mA, 200 mA or 400 mA).

With EDTA at 4oo mA the temperature will be 55 degrees C and you will get
about 80% more calcium removed than at room temperature. Personally I think
it's the raised temperature rather than the alternating current anyway, but
I'm told temperature on its own is inconsistent. It's in Lillie's bible page
794.

Kemlo Rogerson
Pathology Manager
Ext  3311
DD   01934 647057
Mob 07749 754194
 
 
 

-----Original Message-----
From: maria.morell <@t> lab.upc.edu [mailto:maria.morell <@t> lab.upc.edu] 
Sent: Monday, February 20, 2006 1:07 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RDO question

Good morning Histonet!

We are using RDO (a rapid decalcifier with hydrochloric acid) in our
laboratory to decalcify very high density ear bones that surround the
inner ear of marine mammals. The inner ear cells are fixed with formalin
10% and this is the soft tissue we are interested in.

Our problem is that the bone surrounding the inner ear is not homogeneous.
So, while some points are decalcified after 16 hours other
need much more time. Then the inner ear cells are in contact with the RDO
much before the sample is completely decalcified.

I would like to ask if someone know how is the reaction we can expect from
the cells fixed in formalin in contact with RDO.
I'm wondering what you can advise me to do in our case, when the bone is
not homogeneous, but it has to be decalicified.

Thank you very much for your help.

Best regards from,
Maria Morell



Maria Morell
Laboratori d'Aplicacions Bioacústiques
Centre Tecnològic de Vilanova i la Geltrú
Universitat Politècnica de Catalunya
Avda. Rambla Exposició s/n
08800-Vilanova i la Geltrú (Barcelona)- Spain
Telf: 0034 938967227
http://www.lab.upc.edu



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