[Histonet] decalcification of crista-biopsies

[Nick Kirk]

Hans

Probably the best way of determining the end point of decalcification is the
Ammonium hydroxide/ammonium oxalate method as detailed below.

SOLUTIONS:

Solution A, 5% aqueous Ammonium Hydroxide
Solution B, 5% aqueous Ammonium Oxalate

METHOD:

Pipette 5 ml solution A into suitably sized flask or tube.
Add 5 ml of solution B
Add 5 ml of solution from the bottom of decalcification vessel. (Avoid
picking up particulates in the pipette.)
Let stand 15 minutes.
If the test aliquot is clear, decalcification is completed and solution is
still usable.
If cloudy, the solution is exhausted, indicated by the precipitated calcium
oxalate, and decalcification is not complete.
Change the fluid and proceed, testing frequently.

Hope that's of some use.

Nick Kirk
Histopathology
Hinchingbrooke Hospital
Huntingdon
England



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Hans Ooms
Sent: 01 February 2004 10:14
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] decalcification of crista-biopsies


Dear techs, scientists or whatsoever,

The method of decalcifying bony tissue (cristabiopsies, femurheadslices
etc.) I have experience with is by putting the specimen in a 5%
sulphosalicylic acidsolution (or EDTA).
The problem is: "How long should decalcification last to preserve morphology
?". I don't know a method of control. If the biopsie "feels like a sponge"
it was taken out and routine processing (dehydration/alcoholrange,
clearing/xylene and paraffinimpregnation in a VIP) was performed. Nearly
every time morphology was awful !

Crista-biopsies are usually processed by means of MMA-resin (great
morphology!) but this would be to expensive for all bony tissues.

Despite Benno Romeis, Gurr, Junqueira I couldn't find anything satisfactory
!!

Someone out there able to help?

Thanks in advance.

Hans Ooms (HT/The Netherlands).

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