[Histonet] understanding reagents in decalcifier; making it in-house

[Morken, Timothy]

Merissa,

     Water                      77-80	solvent
 Formic acid                 21-23	active ingredient
    Fluorad                       >1		surfactant  - a wetting agent to make the solution wet the bone more easily
 Sodium citrate               >1	emulsifier , buffer
Polyvinyl pyrrolidone      >1	emulsifier 

They say less than one percent of the last three, but you really have no idea whether that is 1%, .1% or .01%. It could be any of those.

But all those surfactants and emulsifiers are meant to keep the solution viable for long periods on the shelf. When you make it fresh you don't really need them.

You can either buy a different decalcifier, or make your own. Making your own with just the water and acid will work just fine. 


Tim Morken
Pathology Site Manager, Parnassus 
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center



-----Original Message-----
From: M.O. via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Wednesday, July 22, 2015 1:24 PM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] understanding reagents in decalcifier; making it in-house

Hello Histonet,

The supplier for our decalcifier, TBD-2 from Shandon, is having issues with getting the product out and we will not be receiving it for at least another month.  Our samples are piling up and I don't know what I should do, but maybe I can make the decalcifier in-house.  I am wondering if I can make my own based on the reagents they listed and their percentages and if certain reagents are not actually necessary.

The samples we typically decalcify are mouse knees (decal time = 2 days), mouse spines (3 days), human bone slabs about 7mm in thickness (7-12 days).  Fixation is in zinc buffered formalin, then decalcification, then 70% EtOH.  Our choice to use TBD-2 is due to the gentle decalcification for IHC and we get GREAT results.

Composition of Shandon TBD-2 Decalcifier:
Component                Weight %
     Water                      77-80
 Formic acid                 21-23
    Fluorad                       >1
 Sodium citrate               >1
Polyvinyl pyrrolidone      >1


If you have any input on what reagents I should use and the percentages for making a decalcifier myself, it would be much appreciated!

Thank you for you help,
Merissa
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