[Histonet] Re: Bird head stored in 70% alcohol and possible decalcification

[Rui TAHARA]

Thank you for helpful suggestions. 
I have further questions. 
Yes, I have a bird head (probably 1 cm X 1 cm ) stored in 70 % ethanol.
But i have a similar size bird head fixed in 3.7% formalin for over night and am actually processing the head to store in 70% ethanol since my lab is just ordering the decalcifying solution. I need to decalcify this sample later. 
But i am wondering if it is better to keep the sample in formalin for a week or so till i get the decalcification solution or i should store it in 70 % ethanol and then fix it for a few days again later? 
I am afraid that longer fixative time would affect the sample somehow (e.g. the sample become too rigid?) 

Thank you, 

rui 

> From: gayle.callis <@t> bresnan.net
> To: histonet <@t> lists.utsouthwestern.edu
> Date: Wed, 4 Mar 2015 16:25:28 -0700
> Subject: [Histonet] Re: Bird head stored in 70% alcohol and possible	decalcification
> 
> You wrote: 
> 
>  
> 
> I have an adult bird skull that fixed with formalin and then has been stored
> in 70% ethanol.  
> 
> I have seen the post that the sample stored in 70% ethanol can be walking
> back through to series of ethanol to water and can be decalcified if it
> needs to be. 
> 
>  
> 
> I am wondering if anybody has done this and there is any side effects from
> decalcification after going through dehydration and rehydration of a sample
> compared to a general straight forward protocol from decalcification to
> dehydration? 
> 
> ****************************************************************************
> ****************************************************************************
> ********************************
> 
>  
> 
> I have, in the past, when a weekend arrive, I interrupted acid bone
> decalcification by removing it from acid decalcifier, a quick water rinse
> and immersed into 70% alcohol before returning bone to fresh acid
> decalcifier the next working day. The bones always decalcified without
> problems but I am sure the decalcification took longer since partially
> decalcified bone had to rehydrate.  I later learned more about dipolar (hope
> I said that correctly) alcohol slowing and/or stopping ionization of calcium
> and ceased using 70% alcohol to interrupt acid decalcification.   I now use
> NBF to interrupt decalcification.  Interestingly, I learned the alcohol
> technique from the AFIP bone pathology lab.     
> 
>  
> 
> Alcohol is put into Perenyi's  nitric acid decalcifying solutions to slow
> down or control very rapid nitric acid decalcification.     
> 
>  
> 
> You did not say how big the bird skull was?   I suggest immersing the skull
> back into NBF to let it totally rehydrate for several days (depending on
> skull size and if the brain is present).  I suggest changing NBF if you
> rehydrate longer than a day.   You don't need to go back through an alcohol
> gradient since many processing schedules have tissue samples  going from NBF
> directly into 70%.   If you leave residual alcohol in the bones, the acid
> decalcification could be slower and hopefully not retarded in any way.
> 
> 
>  
> 
> It certainly is worth a try.   Good luck.
> 
>  
> 
> Gayle M. Callis
> 
> HTL/HT/MT(ASCP) 
> 
>  
> 
>        
> 
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet